Ruthenium complexes for DSSC [26, 27, 28, 29, 30, 31, 32, 33].
\\n\\n
Released this past November, the list is based on data collected from the Web of Science and highlights some of the world’s most influential scientific minds by naming the researchers whose publications over the previous decade have included a high number of Highly Cited Papers placing them among the top 1% most-cited.
\\n\\nWe wish to congratulate all of the researchers named and especially our authors on this amazing accomplishment! We are happy and proud to share in their success!
\\n"}]',published:!0,mainMedia:null},components:[{type:"htmlEditorComponent",content:'IntechOpen is proud to announce that 179 of our authors have made the Clarivate™ Highly Cited Researchers List for 2020, ranking them among the top 1% most-cited.
\n\nThroughout the years, the list has named a total of 252 IntechOpen authors as Highly Cited. Of those researchers, 69 have been featured on the list multiple times.
\n\n\n\nReleased this past November, the list is based on data collected from the Web of Science and highlights some of the world’s most influential scientific minds by naming the researchers whose publications over the previous decade have included a high number of Highly Cited Papers placing them among the top 1% most-cited.
\n\nWe wish to congratulate all of the researchers named and especially our authors on this amazing accomplishment! We are happy and proud to share in their success!
\n'}],latestNews:[{slug:"intechopen-authors-included-in-the-highly-cited-researchers-list-for-2020-20210121",title:"IntechOpen Authors Included in the Highly Cited Researchers List for 2020"},{slug:"intechopen-maintains-position-as-the-world-s-largest-oa-book-publisher-20201218",title:"IntechOpen Maintains Position as the World’s Largest OA Book Publisher"},{slug:"all-intechopen-books-available-on-perlego-20201215",title:"All IntechOpen Books Available on Perlego"},{slug:"oiv-awards-recognizes-intechopen-s-editors-20201127",title:"OIV Awards Recognizes IntechOpen's Editors"},{slug:"intechopen-joins-crossref-s-initiative-for-open-abstracts-i4oa-to-boost-the-discovery-of-research-20201005",title:"IntechOpen joins Crossref's Initiative for Open Abstracts (I4OA) to Boost the Discovery of Research"},{slug:"intechopen-hits-milestone-5-000-open-access-books-published-20200908",title:"IntechOpen hits milestone: 5,000 Open Access books published!"},{slug:"intechopen-books-hosted-on-the-mathworks-book-program-20200819",title:"IntechOpen Books Hosted on the MathWorks Book Program"},{slug:"intechopen-s-chapter-awarded-the-guenther-von-pannewitz-preis-2020-20200715",title:"IntechOpen's Chapter Awarded the Günther-von-Pannewitz-Preis 2020"}]},book:{item:{type:"book",id:"8833",leadTitle:null,fullTitle:"Habitats of the World - Biodiversity and Threats",title:"Habitats of the World",subtitle:"Biodiversity and Threats",reviewType:"peer-reviewed",abstract:"Today it is not easy to talk about habitats and to think about the various threats facing them. We are living in an age in which we are poised between having everything immediately, and maintaining good living conditions on Earth. Unfortunately, this is almost impossible!For this reason it is important that everyone understands the importance of the habitats of the world and the inhabitants: including humans!This book aims to describe some of the world's habitats, their characteristics, and their daily threats. This is done in the hope that our children will see all of this tomorrow. Enjoy reading!",isbn:"978-1-78984-487-0",printIsbn:"978-1-78984-486-3",pdfIsbn:"978-1-83968-008-3",doi:"10.5772/intechopen.81140",price:119,priceEur:129,priceUsd:155,slug:"habitats-of-the-world-biodiversity-and-threats",numberOfPages:158,isOpenForSubmission:!1,isInWos:null,hash:"4b7673e0edb8a67093ee8f925f1c1614",bookSignature:"Carmelo Maria Musarella, Ana Cano Ortiz and Ricardo Quinto Canas",publishedDate:"January 29th 2020",coverURL:"https://cdn.intechopen.com/books/images_new/8833.jpg",numberOfDownloads:2288,numberOfWosCitations:0,numberOfCrossrefCitations:4,numberOfDimensionsCitations:7,hasAltmetrics:1,numberOfTotalCitations:11,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"October 25th 2018",dateEndSecondStepPublish:"December 6th 2018",dateEndThirdStepPublish:"February 4th 2019",dateEndFourthStepPublish:"April 25th 2019",dateEndFifthStepPublish:"June 24th 2019",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6",editedByType:"Edited by",kuFlag:!1,editors:[{id:"276295",title:"Dr.",name:"Carmelo Maria",middleName:null,surname:"Musarella",slug:"carmelo-maria-musarella",fullName:"Carmelo Maria Musarella",profilePictureURL:"https://mts.intechopen.com/storage/users/276295/images/system/276295.jpg",biography:"Carmelo Maria Musarella, PhD (Reggio Calabria, Italy –\n23/01/1975) is a biologist, specializing in plant biology. 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However, there are materials that have complex responses when subjected to stress or strain and present a combination of both responses; such compounds have viscoelastic or viscoplastic responses. The term viscoelasticity is used to describe materials that when subjected to a deformation, have a combination of responses and can flow and at the same time have an elastic response, indicating that such materials can recover from deformation. This type of response in materials is most commonly represented by macromolecular materials. A material which exhibits viscoplastic response, when subjected to stress, can no longer recover its original form. In this book we seek to compile works on materials that exhibit both viscoelastic and viscoplastic responses when subjected to stress or defined strain, and present new methodologies for the evaluation of these types of materials.
",isbn:null,printIsbn:"979-953-307-X-X",pdfIsbn:null,doi:null,price:0,priceEur:null,priceUsd:null,slug:null,numberOfPages:0,isOpenForSubmission:!1,hash:"68c9f6836be98c144b843db45dd48f3c",bookSignature:"Dr. Jose Luis Rivera Armenta and Dr. Beatriz Adriana Salazar-Cruz",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/7675.jpg",keywords:"Viscoelasticity, Viscoelastic Materials, Fractional Viscoelastic Model, Pseudoplastic Fluid, Damping, Viscoelastic Phenomena, Viscoelastic Models, Linear Viscoelasticity, Viscoplasticity, Kernels Model, Finite Element Model, Rheological Test, Viscoleastic Tests, Deformation Stress Tests, Stress Relaxation, Creep Test, Rheology Of Reinforced Composites",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:0,numberOfDimensionsCitations:0,numberOfTotalCitations:0,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"March 11th 2019",dateEndSecondStepPublish:"April 1st 2019",dateEndThirdStepPublish:"May 31st 2019",dateEndFourthStepPublish:"August 19th 2019",dateEndFifthStepPublish:"October 18th 2019",remainingDaysToSecondStep:"2 years",secondStepPassed:!0,currentStepOfPublishingProcess:5,editedByType:null,kuFlag:!1,biosketch:null,coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"107855",title:"Dr.",name:"Jose Luis",middleName:null,surname:"Rivera Armenta",slug:"jose-luis-rivera-armenta",fullName:"Jose Luis Rivera Armenta",profilePictureURL:"https://mts.intechopen.com/storage/users/107855/images/system/107855.jpeg",biography:"José Luis Rivera-Armenta was born in Tampico, Mexico, in 1971. He earned his BSc in Chemical Engineering in 1994, an MSc in Petroleum Technology and Petrochemicals in 1998, and a Ph.D. in Chemical Engineering in 2002 at the Technological Institute of Madero City (ITCM). Since 2003 he has been a full-time professor in postgraduate programs at ITCM and a project manager of several developments sponsored by the National Technologic of Mexico and CONACYT. He has been a member of the National Research System at CONACYT level 1 since 2005. His responsibilities include injection and extrusion and thermal analysis at the laboratory at the Petrochemical Research Center at ITCM. He has advised on nine Ph.D\\'s, 16 master’s degrees, and four bachelor theses and also supervised three post-doctorate students. 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From 1995 to 2008 she gained experience in the chemical process industry (Dynasol Elastomers) developing projects with SBS, SBR and SEBS polymers and characterizing and innovating the quality of products in a wide range of applications: asphalts, adhesives, and compounds. Dr. Salazar-Cruz is the author or coauthor of several scientific publications in English and Spanish, and author or coauthor of several book chapters. She has taught several rheology courses and has also collaborated in several projects supported by the National Technologic of Mexico and CONACYT. 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This requires extensive analysis of developing trends in scientific research in order to offer our readers relevant content. Creating the book catalogue is also based on keeping track of the most read, downloaded and highly cited chapters and books and relaunching similar topics. I am also responsible for consulting with our Scientific Advisors on which book topics to add to our catalogue and sending possible book proposal topics to them for evaluation. Once the catalogue is complete, I contact leading researchers in their respective fields and ask them to become possible Academic Editors for each book project. Once an editor is appointed, I prepare all necessary information required for them to begin their work, as well as guide them through the editorship process. I also assist editors in inviting suitable authors to contribute to a specific book project and each year, I identify and invite exceptional editors to join IntechOpen as Scientific Advisors. 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Churchill, Maja Dutour Sikirić, Božana Čolović and Helga Füredi Milhofer",coverURL:"https://cdn.intechopen.com/books/images_new/8812.jpg",editedByType:"Edited by",editors:[{id:"219335",title:"Dr.",name:"David",surname:"Churchill",slug:"david-churchill",fullName:"David Churchill"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7960",title:"Assorted Dimensional Reconfigurable Materials",subtitle:null,isOpenForSubmission:!1,hash:"bc49969c3a4e2fc8f65d4722cc4d95a5",slug:"assorted-dimensional-reconfigurable-materials",bookSignature:"Rajendra Sukhjadeorao Dongre and Dilip Rankrishna Peshwe",coverURL:"https://cdn.intechopen.com/books/images_new/7960.jpg",editedByType:"Edited by",editors:[{id:"188286",title:"Associate Prof.",name:"Rajendra",surname:"Dongre",slug:"rajendra-dongre",fullName:"Rajendra Dongre"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"72108",title:"Ligands and Coordination Compounds Used as New Photosensitized Materials for the Construction of Solar Cells",doi:"10.5772/intechopen.92268",slug:"ligands-and-coordination-compounds-used-as-new-photosensitized-materials-for-the-construction-of-sol",body:'The current environmental problems and the energy crisis have led to creating new technologies. The renewable energies such as: biofuels, biomass, wind, geothermal, hydraulic, solar, tidal, among others become the main source of the energy generation. The use of solar cells represents an alternative among renewable energies. The development of new materials goes from inorganic structures until polymers, small molecules as organic photovoltaic (OPV) and photosensitized organic materials [1].
In this context, the discovery of ultrafast charge transfer between the semiconductor polymer and inorganic semiconductors have allowed that the OPV have passed in a decade of values close to 1% of efficiency until exceeding 10% [2]. This rapid evolution is motivated by its high potential for generating flexible, lightweight and low cost panels changing the classic concept of panels photovoltaic. In the case of the traditional polymers, the electrons are highly localized and require great energy to be excited (>5 eV) converting them into electrical insulators. In contrast, in conjugated polymer and structures the electrons from Z orbitals form π-type bonds that are associated with lower energies, corresponding to the range of ultraviolet and visible radiation. In molecular solids the transition π → π* that occurs between the occupied molecular orbital of higher energy (HOMO) and the lower energy of orbital (LUMO) determine the equivalent to the forbidden band energy of inorganic semiconductors [3, 4]. On the other hand, they had developed sensitized cell from organic dyes. These are also called Grätzel cells. Photoelectrons that introduce into the conduction band of TiO2 that works as a semiconductor, under light illumination [5].
Dye-sensitized cell (DSSC) have been developed as functional biomimetic models of biological process. In the nature exists dyes with electronic properties that allows to purpose design news in solar panel. Chlorophyll, constituted an example in where there is light absorption and charge-carrier transport. The organic molecule is coupled to semiconductor enhancing the Gap band. This electronic transfer promote absorption to the visible region, which increase its applications [6].
The researchers in a world context have designed, developed and synthetized ruthenium complexes, porphyrins, metal-free organic dyes and organic molecules in this field.
The efficiency of DSSC depends on different requirements listed below [7, 8]:
Broad and strong absorption, preferably extending from energies greater than 900 nm.
The dye needs to be photochemically, thermally, and electrochemically robust within the DSSC in order to withstand the harsh conditions of a practical module.
Firm, irreversible adsorption to the semiconductor’s surface (TiO2) and strong electronic coupling between its excited state and the semiconductor conduction band.
Reduction potential is sufficiently higher than the semiconductor conduction band Edge in order to enable charge injection.
Chemical stability in the ground and the excited states for rapid dye regeneration and charge-injection processes.
Different types of dyes have been tested in the DSSC setting, including: transition-metal complexes, organic dyes, porphyrins and phthalocyanines [9, 10, 11, 12]; however, in terms of photovoltaic performance and long-term stability, Ru(II) complexes comprise the most successful family of DSSCs sensitizers, shown in Table 1. A study on these champion dyes reveals that majority are derivate of N3. The N3 dye represents the first high-performance Ru(II) sensitizer reported in 1993 by Grätzel and co-workers [13], affording power conversion efficiencies of 10.3%. The chemical modification of N3 and N719 is made possible because only two anchoring groups are necessary for electron injection [14, 15] thus various functional groups can be installed to block the electrolyte from interacting with the surface or absorb more light. Then, the performance of these successful sensitizers encompasses ligands that combine extended π-conjugated systems, aspiring to enhance the optical absorptivity of the semiconductor’s surface, along with long hydrophobic alkyl chains, aiming an increase of tolerance against water attack (Table 1). Equally importantly N719, which essentially differ only in the protonation state of tetra-protonated parent dye N3, afford a nearly quantitative conversion of incident photons into electric current over a large spectral range. The improved efficiency of N719, was mainly attributed to the increased cell voltage. Since 1993, chemical modifications of these early Ru(II) complexes have led to researchers achieving power conversion efficiencies up to 11.7% (C106 dye) [16, 17, 18, 19, 20, 21, 22, 23, 24], where one of the DCBPy ligands has been replaced with an extended conjugation using thiophenes and long alkyl chains, lastly, these prevents interfacial recombination [25].
The so-called solar cells sensitized by a dye are a type of hybrid devices that have reached a higher degree of development so far. Within his field, porphyrins represent a very interesting alternative because there is efficient model harnessing sunlight. These systems can be synthetized in bulk heterojunction (BHJ) organic solar cells. The interaction of macrocycles with metal ions such as: Fe2+, Fe3+, Co2+, Co3+, Ni2+, Zn2+, Cu2+, Ru2+, Pd2+ and Pt2+ and hydrogen, alkyl, cycloalkyl, cyclohexyl, cycloheptyl, cyclooctyl, haloalkyl, perhaloalkyl, ether chains have permitted the stabilizations of promising new collections. In dye-sensitized solar porphyrin-based push-pull.
photosensitizers have demonstrated their potential as large and rigid planar conjugated structures, which enhance p-electron delocalization and promote intermolecular π-π interaction, as well charge transport in devices. A problem that they can presents is the effect by lack of light-harvesting beyond 850 nm, thus limiting their cell performance. In the papers, it has been reported that 50% of the total solar phonon flux is located in the red and near-infrared spectra. Zhu and colleagues had reported in 2016 that is quite urgent to develop efficient NIR absorbing molecules for high performance organic solar cells. In the next table, the authors show different publications about the development of new bioinspired porphyrin materials (Table 2).
The DSSC free organic dyes are sensitized molecules whose perspective are aimed at staking, on top of one other in order to obtain panchromatic absorption. Table 3 shows azo, cyano, thiophene, and carbonyl with highly conjugated. A PCE value at 14.7 has been reported by Kakiag et al. [49]. The PCE increase with Voc and Jsc and the best properties were associated with carboxylic group and highly polarizability in the presence of nitrile group.
The extension of the conjugated chain and the substitution of the thiophene groups do not represent a marked difference that allows concluding a relationship between the photovoltaic properties and the structure.
Therefore, this article reports the bibliographic revision for these compounds, specifying the following parameters: Chemical name, abbreviation, structure, power conversion efficiencies (PCE), Jsc (short-circuit-current), Voc (open circuit voltage) electrolyte used and authors.
On the other hand, in this research also have been reported the theoretical studies towards the effect the spacer molecule in macrocycles. The linear molecule designed was a benzothiophene derivate (T) and the spacer selected were o- m or p-diphenyldiamine. The spacer represented the communication channel between linear chains, denominated T. The stabilization of the macrocycles depends of the good assembly. The authors reported a study relationship with the photovoltaic properties for three macrocycles in function of isomeric effect in the spacer. The calculations were performed using Gaussian 09 16-18, program with B3LYP functional [58, 59, 60, 61] and 6-31-6 (d, 2p) as basis set [64] in order to investigate the molecular geometry, electronic structures, and optical properties of o-PDT, m-PDT and p-PDT (Figure 1).
Chemical structure for (a) p-PDT, (b) m-PDT and (c) o-PDT.
The stationary point was estimated with level of theory reported previously for the authors [64]. Finally, the authors through the Lewis acid incorporation showed an electronic improvement mechanism. The acid Lewis effect, as evaluated considering the tetracoordinated mode around metal center (Figure 2).
Geometry optimization for (a) T and (b) TZn.
To explore and understand the electronic and optoelectronic properties of photosensitized materials with application in OPV technology, many theories have emerged. One of the most important and common theories is the theory of functional density (DFT), which is a tool that allowed to establish any property used in photosensitized materials, quantum state of atoms, molecules and solids, making modeling and simulation possible of complex systems with millions of degrees of freedom. At present, DFT has grown tremendously and has become one of the main tools in theoretical physics and molecular chemistry. Modeling in the framework of computational chemistry of photosensitized systems made up of electron donors and electron acceptors ultimately influences photo induced electron transfer and energy reactions. Numerous studies using the Density Functional Theory (DFT) methodology to design, evaluate and predict photovoltaic properties of photoactive materials with application in OPV have been published. The approximation of the theory of functional density (DFT) implemented was Gaussian 09 together with the functional correlation (B3LYP) and the base set 6-31g (d, 2p). This calculation allows optimization of geometry without symmetry restrictions for stationary points. In addition, it provided information on the harmonic frequency analysis, which allows the optimized minimum to be verified. The local minimum is identified when the number of imaginary 32frequencies is equal to zero.
The analysis of the changes in electron density for a given electronic transition was based on the electron density difference maps (EDDMs) constructed using the GaussSum suite of programs. Gásquez and co-workers had proposed two different electronegativities (X) for the charge transfer process: one that describes fractional negative charge donation
Thus, the construction of a so-called donor-acceptor map (DAM) has been suggested. A DAM graphic can be constructed by plotting the values of (y-axis) and
The photovoltaic properties are calculated according to the Scharber model, which is an empirical model for predicting the PCE of organic cell solar. HOMO-LUMO as orbital border under solar irradiation with AM 1.5 G (ASTM G173). The PCE was expressed by the following Eqs. (3) and (4), in where Voc is the open circuit voltage, and Jsc is short circuit current.
where FF is a fill factor of 0.75, Eq. (4):
(q = elementary charge, EQE = external quantum efficiency, ϕ = irradiation flow with AM 1.5 G, and λ = wavelength), and Pinc (incident light power).
On the other hand, the valor corresponding ΔEGAP was calculated as, Eq. (5):
LHE (light capture efficiency determinate), (f = oscillator strength) and Es1 = Excitation energy for λmax, Eq. (6).
The excitation energies (Es1) presented in Table 4 were relatively small for the PDT molecule ligand, which indicate a shift to visible region in relationship with λmax. The p-PDT showed the lowest value for Es1, which is directly correlated with the conversion energy (PCE). On the other hand, the cycle size generated for o-PDT and m-PDT systems is smaller, but this does not guarantee a better transfer. On the contrary, there is less efficient in the electronic transport.
Molecule | Wavelength λmax | Es1 (eV) | F | LHE |
---|---|---|---|---|
o-PDT | 432.68 | 2.874 | 2.0217 | 0.9905 |
m-PDT | 425.37 | 2.923 | 2.8755 | 0.9987 |
p-PDT | 465 | 2.67 | 0.07 | 0.14 |
Optical properties for (a) o-PDT, (b) m-PDT and (c) p-PDT.
The LHE values were 2.0217, 2.8755 and 0.07 for o-PDT, m-PDT and p-PDT, respectively. This indicates that o-PDT and m-PDT had a similar sensitivity to sunlight and will reflect higher values of LHE compared to p-PDT.
The visible light absorption ability may benefit from absorbing more photons and generating high photocurrent, which is a strong advantage of T derivatives. In the previous reports, PD spacers that cannot absorb visible light were observed. It is necessary that T derivate linked to the PD fragment enhances the electronic coupling in the excited state, which operates as a gated wire in π-conjugated systems, as is observed for o-PDT, m-PDT and p-PDT (Figure 3). The isomeric effect is greatly correlated to geometric distortion o-PDT and m-PDT molecules, which were dramatically affected in relationship to its planarity. The cavity between linear molecules is small, but the torsion affects the electronic properties.
Theoretical spectra electronic for (a) o-PDT m-PDT.
The effect of Lewis acid on macrocycle stabilization is shown below. The geometric environment of the metallic center was tetrahedral, considering two positions to the electro donator atoms corresponding to linear and macrocycle molecule; and two water molecules. The incorporation of the metal into the linear chain (ZnT) generates a decrease in the value for GAP around 1.72 eV, a value located in the visible region. However, the effect is more severe when incorporated into the macrocycle, in where; its addition generated a decrease in GAP still 1.55 eV (p-ZnPDT). The DAM graphic for these systems indicated a significant improvement in donor capacity. These criteria are important to electronically activate the photovoltaic cell (Figure 4).
DAM graphic for T, p-PDT, ZnT and p-ZnPDT.
The spectra in Figure 5 showed a similar profile for TZn, and p-ZnPDT with the incorporation of Lewis acid in the structure, which have an intense main band to 568 nm, and 516 nm respectively. This band corresponds to the dominant electron transition from HOMO to LUMO, that is, from the π molecular orbital (chromophore fragments-π-linker) to the π* orbital (acceptor fragment), and this process can be ascribed to the intramolecular charge transfer.
Theoretical spectra electronic for TZn and p-ZnPDT.
The results showed in Table 5 suggested decreased the ΔEGAP in relationship with PCE. These results are congruent with the optical, and electronic properties observed previously. The p-PDT presented the best photovoltaic properties. The metal ion generates a symmetrical tension in the system, and this could explain its behavior. The Jsc increased in function of decreased the ΔEGAP, concluding that the preferential isomer for the construction of this family macrocycles is the p-PDT, considering theoretical models in the gas phase.
Molecule | ΔEGAP (eV) | Jsc (mA/cm2) | Voc (V) | FF | PCE (%) |
---|---|---|---|---|---|
T | 3.07 | 5.36 | 2.77 | 0.75 | 11.12 |
p-PDT | 2.66 | 14.79 | 2.36 | 0.75 | 26.18 |
ZnT | 1.72 | 21.52 | 1.42 | 0.75 | 22.92 |
p-ZnPDT | 1.55 | 11.76 | 1.25 | 0.75 | 11.02 |
Photovoltaic parameters for T, p-PDT, ZnT and p-ZnPDT.
The purpose of this review of DSSC materials was to compile the information reported to: synthetized ruthenium complexes, porphyrins, and metal-free organic dyes. For researchers, it is important to know parameters such as: PCE, Jsc, and Voc; which help you to diffuse between structures, and propose synthesis strategies that make possible new materials in this field application. Principles for the future development of new molecules can be analyzed and likewise it is interesting support to follow up structure families as a function of time. Although many structures are shown here, there is still a need to optimize the chemical, and physical properties to promote improved solar cells. On the other hand, in this work, the best photovoltaic parameters were described for p-PDT with PCE 26.18%, Jsc = 14.79 mA/cm2, and ΔE = 2.66 eV such as macrocycle. The metal ion influences the electronic properties, and decreases the ΔEGAP. The incorporation of Lewis acid in the structure macrocycle to increase of the optical properties, which allows rigidity that can benefit planarity.
This work was supported by Universidad Santiago de Cali—DGI Grants 63661. The author acknowledgment to Melissa Suarez for technical support and Hoover Valencia for data acquisition.
The authors declare that they have no conflict of interests.
Maize (Zea mays L.) is an important staple food crop in sub-Saharan Africa (SSA). It is the third most important cereal crop after wheat and rice [1]. It is used for both livestock feeds and human consumption. In SSA, maize accounts for about 70% of the human food [2]. The demand for maize is expected to increase by >90.0% in SSA by 2020 [3]. However, the productivity of the crop is limited by several abiotic and biotic stresses. Among these abiotic factors, insect pests, such as the stem borers and weevils, cause considerable economic damage on the crop [4, 5]. In addition, fungal diseases such as gray leaf spot (Cercospora zeae-maydis Tehon & Daniels), common leaf rust (Puccinia sorghi Schr.), and turcicum leaf blight (TLB) (Exserohilum turcicum) often pose a serious threat to maize production [6].
\nIn particular, TLB, also known as the northern corn leaf blight, can devastate the crop in high rainfall, humid areas [6, 7]. TLB reduces the seed quality, resulting in diminished germination capacity, low sugar content as well as predisposition to stalk rot [8, 9]. The use of resistant varieties is an inexpensive method for combating TLB [10]. Currently, there are efforts to incorporate durable resistance into maize germplasm particularly in SSA where some commercial varieties as well as elite parental inbred lines are reportedly vulnerable to TLB [11, 12]. For example, in Ethiopia, maize productivity is low (averaging about 2.5 t/ha) in the smallholder production systems partly due to TLB and other stresses. Spurred by the need to enhance maize productivity for farmers, the national maize improvement program in Ethiopia recently embarked on a breeding project aimed at developing leaf blight resistant hybrid varieties that are adapted to the major maize-growing areas of the country which are predominantly in the mid-altitude to subhumid agroecologies [13]. However, hybrid breeding for resistance to leaf blight requires knowledge of the genetic variability of the germplasm in terms of its reaction to TLB as well as its characterization into distinct genetic groups that can be hybridized in order to exploit heterosis.
\nThe variability in the host (maize) plant resistance to the disease occurs in either the qualitative or the quantitative form. The qualitative form of resistance is race specific and is governed by a single or few genes but the quantitative form of resistance is race nonspecific and polygenic [14, 15]. In addition, qualitative resistance can break down due to the emergence of new virulent races of the pathogen through genetic mutation and recombination events [12, 15]. The pathogen E. turcicum exhibits a wide range of variability [16], and new races are capable of overcoming previously resistant varieties [7]. For instance, the resistance conferred by the Htn gene(s) is characterized by chlorotic and necrotic lesions or lesions surrounded by a yellow-to-light-brown margin (without spore formation), which limits the growth and spread of the disease [12, 14]. In contrast, the resistance conferred by Htn gene is expressed as a delay in lesion formation typically showing at the pollination stage [17, 18]. Lesion size, together with area under disease progress curve (AUDPC) as well as disease severity and incidence, are commonly used in evaluating maize genotypes for resistance to TLB [19, 20]. However, phenotypic evaluations in conventional breeding approaches are unable to detect the presence of favorable alleles in the germplasm. Therefore, marker-assisted selection and DNA fingerprinting techniques have been effectively used to increase the efficiency of conventional breeding, particularly the time required for developing new improved varieties in maize [12].
\nThe presence of discrete genetic groups among inbred lines is attributed to increased allelic diversity which is useful in optimizing hybrid vigor. Assigning inbred lines into well-differentiated genetic clusters can reduce the creation and evaluation of many undesirable crosses [21]. Molecular markers assist in characterizing inbred lines and in establishing distinct clusters of genotypes based on genetic diversity, which is useful in maize breeding programs [22, 23]. Molecular markers were applied successfully to allocate maize germplasm into heterotic groups [24–26]. In a study which compared different markers for their effectiveness in estimating genetic grouping among maize inbred lines, SSR markers revealed the highest level of polymorphism due to their codominant nature and high number of alleles per locus [27]. Therefore, the study reported in this chapter was designed to evaluate the (i) genetic variability in reaction to TLB among maize inbred lines under field conditions and (ii) diversity of selected medium to late maturity tropical maize inbred lines for hybrid breeding using selected SSR markers.
\nFifty inbred lines were used in the study. The lines were adapted to the mid-altitude agroecologies in Ethiopia and were obtained from the national maize research program and the international maize and wheat improvement center (CIMMYT). Inbred line CML-197, which was obtained from CIMMYT, served as susceptible check (\nTable 1\n). The field trial was conducted at Bako (37°09′ E; 09°06′ N; 1650 m above sea level). It receives approximately 1200 mm rainfall annually (\nTable 2\n) and is representative of the mid-altitude subhumid agroecological region in Ethiopia.
\nEntry | \nPedigree | \nOrigin | \n
---|---|---|
1 | \nCML 202 | \nCIMMYT | \n
2 | \nCML 442 | \nCIMMYT | \n
3 | \nCML 312 | \nCIMMYT | \n
4 | \nCML 464 | \nCIMMYT | \n
5 | \nGibe-1-91-1-1-1-1 | \nBAKO | \n
6 | \nCML 445 | \nCIMMYT | \n
7 | \nCML 443 | \nCIMMYT | \n
8 | \nCML 197 | \nCIMMYT | \n
9 | \nA-7033 | \nBAKO | \n
10 | \nCML 205/208//202-X-2-1-2-B-B-B | \nBAKO | \n
11 | \nCML 395 | \nCIMMYT | \n
12 | \nF-7215 | \nBAKO | \n
13 | \nDE-78-Z-126-3-5-5-1-1 | \nBAKO | \n
14 | \n30H83-7-1-1-1-2-1 | \nBAKO | \n
15 | \nI100E-1-9-1-1-1-1-1 | \nBAKO | \n
16 | \nSZYNA99F2-81-4-3-1 | \nBAKO | \n
17 | \nX1264DW-1-2-1-1-1 | \nBAKO | \n
18 | \n124-b (113) | \nBAKO | \n
19 | \nSC22 | \nBAKO | \n
20 | \nSC715-121-1-3 | \nBAKO | \n
The pedigree and origin of maize inbred lines that were evaluated for diversity using SSR markers.
Month | \n2011 | \n2012 | \n||||
---|---|---|---|---|---|---|
Rainfall (mm) | \nTemperature (C0) | \nRH (%) | \nRainfall (mm) | \nTemperature (C0) | \nRH (%) | \n|
January | \n15.90 | \n20.20 | \n58.00 | \n0.00 | \n20.40 | \n52.70 | \n
February | \n2.00 | \n20.90 | \n50.90 | \n4.40 | \n21.80 | \n47.50 | \n
March | \n58.80 | \n21.90 | \n53.90 | \n16.20 | \n23.00 | \n48.90 | \n
April | \n68.10 | \n20.40 | \n52.40 | \n30.70 | \n24.00 | \n62.50 | \n
May | \n222.20 | \n21.30 | \n58.50 | \n92.8 | \n23.00 | \n55.60 | \n
June | \n295.00 | \n19.90 | \n67.50 | \n153.30 | \n20.20 | \n66.90 | \n
July | \n224.10 | \n19.30 | \n69.30 | \n138.20 | \n19.50 | \n76.00 | \n
August | \n294.60 | \n19.10 | \n75.60 | \n263.60 | \n19.70 | \n64.00 | \n
September | \n131.30 | \n20.00 | \n65.90 | \n157.50 | \n20.10 | \n74.40 | \n
October | \n53.20 | \n20.20 | \n59.80 | \n6.00 | \n21.00 | \n50.50 | \n
November | \n60.10 | \n20.00 | \n59.80 | \n17.10 | \n20.30 | \n49.70 | \n
December | \n0.00 | \n19.80 | \n54.50 | \n6.70 | \n21.5 | \n45.70 | \n
Total | \n1425.30 | \n886.50 | \n
Average monthly rainfall, temperature, and relative humidity at Bako during the 2011 and 2012 cropping seasons.
RH = relative humidity.
Inbred lines were evaluated using the lattice design with three replications. Trials were conducted for two consecutive seasons (in 2011 and 2012) during the main rainy season (May to September) in Ethiopia. The seed of each genotype was planted manually in the field in a two-row plot 5.1 m long × 0.75 m at 30.0 cm intra-row spacing. Phosphorus (in the form of diammonium phosphate) was applied once at planting at 100.0 kg/ha. Nitrogen fertilizer (in the form of urea) was applied at 100.0 kg/ha in two splits with 50% at planting and the rest at 37 days after emergence. Standard maize trial management practices were applied throughout each season at the location.
\nIsolates of E. turcicum were obtained from diseased maize leaf samples that were collected from fields where the disease is prevalent. The infected leaves were excised into small sections (approx. 1.0 cm2 each) prior to surface sterilization using 2.5% Sodium hypochlorite for about 3 min and subsequently rinsed with sterile distilled water and blot-dried before plating on PDA in petri dishes for incubation at room temperature for 3–4 days. Pure cultures were prepared by subculturing from the isolation plates followed by incubation for 7–10 days in order to obtain sufficient growth. The inoculum was prepared by flooding the cultures with sterile distilled water and scrapping the surface with microscopic slides to dislodge the conidia and then filtered using cheese cloth after which the concentration of the conidia suspension was adjusted to approximately 105 conidia per milliliter using a hemocytometer [28].
\nMaize plants growing in the field were inoculated at the four to six leaf growth stages during the middle of the main rainy season (mid-July) in Ethiopia. The inoculations were accomplished by spraying (manually, with the aid of an atomizer) the maize plant with the conidia suspension until runoff after which fine mist water was sprayed over the inoculated plants in order to create conducive conditions for disease development. This inoculation procedure was carried out during the evening when there was sufficient moisture in the air.
\nIn each season, the disease was visually assessed in the field 2–3 weeks after inoculation. Ten randomly selected plants were tagged and used for successive disease assessments. Plants were rated at 10-day intervals for percent incidence, lesion length, and lesion width. In order to determine the rate of lesion expansion, 2 lesions out of the 10 plants were measured (and marked for subsequent tracing) at 10-day intervals.
\nDisease severity was scored using a scale of 1–5 where:
\n1.0 = very slightly infected, one or two restricted lesions on lower leaves or trace.
\n2.0 = slight-to-moderate infection on lower leaves, a few scattered lesions on lower leaves.
\n3.0 = abundant lesions on lower leaves, a few on middle leaves.
\n4.0 = abundant lesions on lower and middle leaves extending to upper leaves.
\n5.0 = abundant lesions on all leaves, plant may be prematurely killed by blight.
\nThe AUDPC was determined from the disease severity scores obtained in both seasons. The AUDPC parameter was calculated using Eq. (1) below as described previously [29]:
\nwhere n = number of observations, ti = number of days after planting for the ith disease assessment, and yi = disease severity.
\nThe parameter was used to quantify the epidemic from the beginning to the peak of the disease. The grain yield was calculated using the average shelling percentage of 80% adjusted to 12.5% moisture. Data sets of the quantitative measurements from individual trials were subjected to standard analysis of variance procedures using the GenStat release 14.2 computer software program [30].
\nTwenty maize inbred lines were used in the study. Eight of these inbred lines were originally developed for the mid-altitude and subhumid agroecologies at CIMMYT, whereas the remainder was developed by the local Ethiopian maize research program and was well adapted to mid-altitude areas. The local inbred lines were developed from three heterotic groups (that are commonly used in the country) namely Kitale synthetic II, Ecuador 573, and Pool 9A.
\nDNA was collected from 3- to 4-week-old plants (tagged for identification), using Whatman FTA cards and the modified protocol of FTA paper technology [31]. Ten DNA samples from each of the 20 inbred lines were then bulked (in order to eliminate variation within each entry) and used for the diversity analysis at the INCOTEC-PROTEIOS laboratory in South Africa (Incotec, SA Pty. Ltd., South Africa) utilizing 20 SSR markers. PCR products of all of the 20 primers were fluorescently labeled and separated by capillary electrophoresis on an ABI 3130 automatic sequencer (Applied Biosystems, Johannesburg, South Africa). Analysis was performed using GeneMapper 4.1. The data matrices of the genetic distances were used to create the dendrogram using the unweighted pair group method with arithmetic mean allocated (UPGMA). The polymorphism information content (PIC) was calculated as:
\nPIC = 1 − ∑fi.
\nwhere fi is the frequency of the ith allele [32].
\nDisease ratings were significantly different among the 50 inbred lines (P < 0.001), and 11 were classified as resistant, 26 as intermediate, whereas the remainder was classified as susceptible (\nTables 3\n and \n4\n). The resistant inbred lines (e.g., 136-a and 142-1-e) attained lower disease severity scores compared to the susceptible check CML-197 (\nTables 3\n and \n4\n). No accession was immune to the disease. In addition, there were highly significant (P < 0.001) differences for lesion length among inbred lines in both 2011 and 2012. The inbred lines Pool9A-4-4-1-1-1, SZSYNA-99-F2-803-4-1, and CML 197 showed comparatively larger lesion lengths, whereas the lesion length of CML 202 and CML 312 showed consistently small lesion lengths over the two seasons. Resistance to E. turcicum in maize germplasm was previously associated with a reduction in percent leaf area as well as small lesions [33].
\nNo. | \nInbred line | \nDSS | \nReaction type | \nIncidence (%) | \nLesion length (cm) | \nAUDPC | \nTSW | \nYield (t ha−1) | \n
---|---|---|---|---|---|---|---|---|
1 | \nCML 202 | \n2.00 | \nR | \n46.81 | \n9.88 | \n408.3 | \n223.3 | \n2.22 | \n
2 | \nCML442 | \n2.734 | \nI | \n78.43 | \n13.40 | \n612.5 | \n223.3 | \n2.40 | \n
3 | \nCML 312 | \n2.413 | \nI | \n61.52 | \n10.35 | \n385.0 | \n276.7 | \n3.03 | \n
4 | \nCML 464 | \n2.210 | \nI | \n55.64 | \n13.82 | \n595.0 | \n223.3 | \n3.79 | \n
5 | \nGibe-1-91-1-1-1-1 | \n2.534 | \nI | \n71.32 | \n14.57 | \n408.3 | \n321.7 | \n2.90 | \n
6 | \nCML 445 | \n2.523 | \nI | \n65.20 | \n14.02 | \n571.7 | \n213.3 | \n3.34 | \n
7 | \nCML 443 | \n2.934 | \nS | \n69.61 | \n13.48 | \n595.0 | \n211.7 | \n2.07 | \n
8 | \nGibe-1-158-1-1-1-1 | \n2.496 | \nI | \n66.42 | \n11.37 | \n507.5 | \n281.7 | \n3.43 | \n
9 | \nA7033 | \n2.881 | \nS | \n68.63 | \n15.37 | \n641.7 | \n273.3 | \n2.58 | \n
10 | \n(CML 205/CML208//CML 202) -X2-1-2-B-B-B | \n2.696 | \nS | \n83.58 | \n15.88 | \n571.7 | \n300.0 | \n5.60 | \n
11 | \nCML395 | \n2.388 | \nI | \n71.08 | \n14.07 | \n420.0 | \n338.3 | \n4.96 | \n
12 | \nCML 444 | \n2.526 | \nI | \n69.12 | \n18.28 | \n443.3 | \n260.0 | \n2.95 | \n
13 | \nDE-78-Z-126-3-2-2-1-1 | \n2.688 | \nS | \n67.89 | \n14.48 | \n536.7 | \n280.0 | \n4.14 | \n
14 | \n30H83-7-1-1-1-2-1 | \n2.00 | \nR | \n53.19 | \n10.90 | \n495.8 | \n210.0 | \n3.14 | \n
15 | \nILoo’E-1-9-1-1-1-1-1 | \n2.00 | \nR | \n56.62 | \n15.62 | \n420.0 | \n346.7 | \n4.83 | \n
16 | \nSZSYNA-99-F2-814-3-1 | \n2.00 | \nR | \n42.40 | \n10.77 | \n466.7 | \n315.0 | \n2.46 | \n
17 | \nX1264DW-1-2-1-1-1-1 | \n2.889 | \nS | \n70.59 | \n15.00 | \n571.7 | \n213.3 | \n1.94 | \n
18 | \n124-b(113) | \n2.559 | \nI | \n59.80 | \n15.27 | \n606.7 | \n365.0 | \n3.53 | \n
19 | \nSC22 | \n2.760 | \nS | \n85.78 | \n14.72 | \n501.7 | \n271.7 | \n3.56 | \n
20 | \nSC-715-1211-3 | \n2.466 | \nI | \n67.40 | \n13.47 | \n396.7 | \n336.7 | \n3.45 | \n
21 | \nDE-105-Z-126-30-1-2-2-1 | \n2.00 | \nR | \n61.27 | \n14.55 | \n420.0 | \n235.0 | \n2.89 | \n
22 | \nGibe-1-20-2-2-1-1 | \n2.663 | \nS | \n69.12 | \n18.78 | \n501.7 | \n301.7 | \n2.62 | \n
23 | \nKuleni-0080-4-2-1-1-1-1 | \n2.022 | \nI | \n61.52 | \n16.38 | \n449.2 | \n326.7 | \n3.72 | \n
24 | \nPool9A-4-4-1-1-1 | \n2.677 | \nS | \n68.63 | \n21.35 | \n670.8 | \n288.3 | \n4.85 | \n
25 | \n30H83-5-1-4-2-1-1 | \n2.486 | \nI | \n63.97 | \n16.27 | \n484.2 | \n308.3 | \n4.27 | \n
26 | \nIloo’E-5-5-3-1 | \n2.639 | \nI | \n74.26 | \n13.48 | \n560.0 | \n328.3 | \n4.41 | \n
27 | \nSZSYNA-99-F2-2-7-3-1-1 | \n2.00 | \nR | \n57.35 | \n11.77 | \n478.3 | \n206.7 | \n2.77 | \n
28 | \nSC-715-154-1-1 | \n2.206 | \nI | \n65.20 | \n11.97 | \n402.5 | \n280.0 | \n5.89 | \n
29 | \nBH6609(F2)-10-2-1-2-1 | \n2.333 | \nI | \n61.76 | \n11.83 | \n402.5 | \n300.0 | \n3.98 | \n
30 | \n143-5-I | \n2.305 | \nI | \n60.29 | \n15.48 | \n420.0 | \n325.0 | \n6.84 | \n
31 | \n144-7-b | \n1.90 | \nR | \n58.09 | \n12.87 | \n385.0 | \n330.0 | \n4.45 | \n
32 | \n(LZ-955459/LZ955357)-B-1-B-B | \n2.369 | \nI | \n67.16 | \n12.20 | \n431.7 | \n256.7 | \n2.98 | \n
33 | \n139-5-j | \n2.00 | \nR | \n53.43 | \n13.78 | \n385.0 | \n258.3 | \n2.56 | \n
34 | \n30H83-56-1-1-1-1-1 | \n2.351 | \nI | \n57.35 | \n10.22 | \n495.8 | \n205.0 | \n3.57 | \n
35 | \nSZSYNA-99-F2-80-3-4-1 | \n2.653 | \nI | \n73.53 | \n20.05 | \n525.0 | \n293.3 | \n3.15 | \n
36 | \n124-b(109) | \n2.901 | \nS | \n81.86 | \n15.48 | \n536.7 | \n310.0 | \n5.54 | \n
37 | \nF7215 | \n2.417 | \nI | \n63.73 | \n14.72 | \n455.0 | \n393.3 | \n3.86 | \n
38 | \n136-a | \n1.80 | \nR | \n51.47 | \n13.82 | \n238.0 | \n396.7 | \n4.41 | \n
39 | \nDE-78-Z-126-3-2-1-2-1 | \n2.631 | \nI | \n70.83 | \n14.85 | \n595.0 | \n286.7 | \n3.83 | \n
40 | \nGibe-1-186-2-2-1 | \n2.549 | \nI | \n51.96 | \n14.88 | \n350.0 | \n373.3 | \n2.70 | \n
41 | \nPool9A-128-5-1-1-1 | \n2.718 | \nI | \n71.43 | \n13.12 | \n595.0 | \n278.3 | \n2.45 | \n
42 | \n30H83-7-1-5-1-1-1-1 | \n2.00 | \nR | \n52.45 | \n12.05 | \n379.2 | \n220.0 | \n2.60 | \n
43 | \nSZSYNA-99-F2-3-6-2-1 | \n2.587 | \nI | \n70.83 | \n12.33 | \n618.3 | \n256.7 | \n2.36 | \n
44 | \nSC-715-13-2-1 | \n2.434 | \nI | \n61.76 | \n12.87 | \n420.0 | \n248.3 | \n2.34 | \n
45 | \nSC-22-430(63) | \n3.033 | \nS | \n80.15 | \n11.57 | \n478.3 | \n311.7 | \n2.48 | \n
46 | \nKuleni-C1-101-1-1-1 | \n3.028 | \nS | \n75.49 | \n17.07 | \n700.0 | \n258.3 | \n2.84 | \n
47 | \nIloo’E-1-12-4-1-1 | \n2.355 | \nI | \n51.96 | \n10.30 | \n443.3 | \n276.7 | \n2.43 | \n
48 | \n(DRB-F2-60-1-2)-B-1-B-B-B | \n2.791 | \nS | \n75.98 | \n16.23 | \n600.8 | \n270.0 | \n2.66 | \n
49 | \n142-1-e | \n2.00 | \nR | \n62.01 | \n15.02 | \n595.0 | \n323.3 | \n3.94 | \n
50 | \nCML 197 | \n3.028 | \nS | \n88.48 | \n18.07 | \n525.0 | \n271.7 | \n50 | \n
\n | LSD | \n0.4260 | \n— | \n18.513 | \n7.504 | \n129.93 | \n72.64 | \n1.465 | \n
\n | Pr > f | \n** | \n— | \n** | \n** | \n** | \n** | \n** | \n
\n | CV (%) | \n3.3 | \n— | \n17.6 | \n10.6 | \n16.2 | \n15.9 | \n25 | \n
\n | Overall mean | \n2.486 | \n— | \n65.49 | \n14.16 | \n493.9 | \n284.1 | \n3.52 | \n
Maize leaf blight reactions, grain yield, and thousand seed weight of 50 inbred lines tested during 2011 at Bako research Center in Ethiopia.
DSS = disease severity score (0.00–5.00); R = resistant (1.0–2.00); I = intermediate (2.10–2.50); susceptible (2.51–5.00); and TSW = thousand seed weight.
** = Significant at 0.05 and 0.01 probability levels, respectively.
No. | \nInbred line | \nDSS | \nReaction type | \nIncidence (%) | \nLesion length (cm) | \nLesion width (cm) | \nTSW | \nYield (t/ha) | \n
---|---|---|---|---|---|---|---|---|
1 | \nCML 202 | \n2.39 | \nR | \n40.69 | \n12.00 | \n1.33 | \n173 | \n2.15 | \n
2 | \nCML442 | \n2.69 | \nS | \n72.55 | \n13.67 | \n1.67 | \n210 | \n2.67 | \n
3 | \nCML 312 | \n2.47 | \nI | \n64.22 | \n12.33 | \n0.83 | \n220 | \n3.25 | \n
4 | \nCML 464 | \n1.92 | \nR | \n52.45 | \n13.00 | \n1.03 | \n207 | \n3.01 | \n
5 | \nGibe-1-91-1-1-1-1 | \n2.53 | \nS | \n74.02 | \n20.33 | \n1.50 | \n260 | \n3.76 | \n
6 | \nCML 445 | \n2.42 | \nI | \n65.69 | \n14.33 | \n1.17 | \n207 | \n3.36 | \n
7 | \nCML 443 | \n2.97 | \nS | \n64.71 | \n13.00 | \n1.00 | \n183 | \n1.93 | \n
8 | \nGibe-1-158-1-1-1-1 | \n2.39 | \nI | \n58.82 | \n12.00 | \n1.57 | \n270 | \n2.93 | \n
9 | \nA7033 | \n2.81 | \nS | \n58.82 | \n13.33 | \n1.33 | \n240 | \n2.41 | \n
10 | \n(CML 205/CML208//CML 202) -X2-1-2-B-B-B | \n2.64 | \nS | \n86.76 | \n22.33 | \n1.83 | \n237 | \n5.83 | \n
11 | \nCML395 | \n2.33 | \nI | \n70.59 | \n21.67 | \n2.00 | \n313 | \n5.04 | \n
12 | \nCML 444 | \n2.61 | \nS | \n65.69 | \n23.33 | \n2.00 | \n230 | \n2.67 | \n
13 | \nDE-78-Z-126-3-2-2-1-1 | \n2.67 | \nS | \n65.20 | \n18.33 | \n1.50 | \n250 | \n2.6 | \n
14 | \n30H83-71-1-1-2-1 | \n1.89 | \nR | \n39.71 | \n13.33 | \n1.67 | \n187 | \n2.91 | \n
15 | \nILoo’E-1-9-1-1-1-1-1 | \n2.14 | \nI | \n54.41 | \n23.67 | \n1.33 | \n293 | \n4.62 | \n
16 | \nSZSYNA-99-F2-814-3-1 | \n1.69 | \nR | \n27.94 | \n14.00 | \n1.00 | \n257 | \n2.01 | \n
17 | \nX1264DW-1-2-1-1-1-1 | \n2.36 | \nI | \n72.55 | \n19.33 | \n1.33 | \n183 | \n1.92 | \n
18 | \n124-b(113) | \n2.34 | \nI | \n45.10 | \n16.33 | \n1.67 | \n303 | \n3.13 | \n
19 | \nSC22 | \n2.05 | \nI | \n91.18 | \n16.67 | \n2.00 | \n230 | \n3.14 | \n
20 | \nSC-715-121-1-3 | \n3.07 | \nS | \n70.10 | \n16.00 | \n2.17 | \n270 | \n2.64 | \n
21 | \nDE-105-Z-126-30-1-2-2-1 | \n1.57 | \nR | \n69.61 | \n20.67 | \n1.83 | \n230 | \n3.42 | \n
22 | \nGibe-1-20-2-2-1-1 | \n2.48 | \nI | \n77.45 | \n25.00 | \n1.33 | \n287 | \n3.08 | \n
23 | \nKuleni-0080-4-2-1-1-1-1 | \n4.29 | \nI | \n58.33 | \n20.33 | \n1.33 | \n283 | \n3.8 | \n
24 | \nPool9A-4-4-1-1-1 | \n2.42 | \nI | \n62.25 | \n25.67 | \n1.67 | \n270 | \n5.35 | \n
25 | \n30H83-51-4-2-1-1 | \n2.47 | \nI | \n67.16 | \n22.00 | \n2.00 | \n260 | \n4.39 | \n
26 | \nIloo’E-5-5-3-1 | \n2.61 | \nS | \n77.94 | \n14.33 | \n1.00 | \n260 | \n3.5 | \n
27 | \nSZSYNA-99-F2-2-7-3-1-1 | \n2.22 | \nI | \n55.88 | \n15.00 | \n1.50 | \n170 | \n2.88 | \n
28 | \nSC-715-154-1-1 | \n2.14 | \nI | \n73.53 | \n15.67 | \n1.83 | \n217 | \n5.01 | \n
29 | \nBH6609(F2)-10-2-1-2-1 | \n2.33 | \nI | \n54.90 | \n15.33 | \n1.53 | \n243 | \n1.74 | \n
30 | \n143-5-I | \n2.08 | \nI | \n51.96 | \n18.00 | \n2.17 | \n273 | \n5.95 | \n
31 | \n144-7-b | \n1.89 | \nR | \n59.31 | \n18.00 | \n1.00 | \n333 | \n3.47 | \n
32 | \n(LZ-955459/LZ955357)-B-1-B-B | \n2.28 | \nI | \n67.65 | \n16.67 | \n1.33 | \n200 | \n2.72 | \n
33 | \n139-5-j | \n2.03 | \nI | \n44.12 | \n19.33 | \n1.07 | \n237 | \n1.8 | \n
34 | \n30H83-561-1-1-1-1 | \n2.22 | \nI | \n50.98 | \n13.00 | \n0.83 | \n203 | \n2.93 | \n
35 | \nSZSYNA-99-F2-80-3-4-1 | \n2.81 | \nS | \n76.47 | \n27.67 | \n1.83 | \n237 | \n3.38 | \n
36 | \n124-b(109) | \n3.03 | \nS | \n82.84 | \n19.33 | \n1.33 | \n270 | \n5.59 | \n
37 | \nF7215 | \n2.5 | \nI | \n62.75 | \n21.33 | \n1.07 | \n273 | \n2.92 | \n
38 | \n136-a | \n1.75 | \nR | \n42.16 | \n17.33 | \n1.33 | \n363 | \n3.62 | \n
39 | \nDE-78-Z-126-3-2-1-2-1 | \n2.56 | \nS | \n65.20 | \n19.00 | \n1.00 | \n237 | \n3.54 | \n
40 | \nGibe-1-186-2-2-1 | \n2.81 | \nS | \n49.02 | \n19.33 | \n1.33 | \n360 | \n2.3 | \n
41 | \nPool9A-128-5-1-1-1 | \n2.67 | \nS | \n68.36 | \n15.33 | \n1.67 | \n223 | \n2.87 | \n
42 | \n30H83-71-5-1-1-1-1 | \n1.94 | \nR | \n50.00 | \n16.67 | \n2.00 | \n193 | \n2.84 | \n
43 | \nSZSYNA-99-F2-3-6-2-1 | \n2.28 | \nI | \n63.24 | \n15.33 | \n2.00 | \n233 | \n2.36 | \n
44 | \nSC-715-13-2-1 | \n2.47 | \nI | \n66.67 | \n16.33 | \n1.17 | \n210 | \n2.34 | \n
45 | \nSC-22-430(63) | \n3.08 | \nS | \n89.71 | \n14.33 | \n1.67 | \n227 | \n2.05 | \n
46 | \nKuleni-C1-101-1-1-1 | \n2.81 | \nS | \n68.63 | \n7.67 | \n1.17 | \n237 | \n3.07 | \n
47 | \nIloo’E-1-12-4-1-1 | \n2.17 | \nI | \n33.33 | \n10.67 | \n1.33 | \n243 | \n1.57 | \n
48 | \n(DRB-F2-60-1-2)-B-1-B-B-B | \n2.72 | \nS | \n67.65 | \n21.00 | \n2.00 | \n230 | \n2.48 | \n
49 | \n142-1-e | \n1.81 | \nR | \n49.51 | \n16.33 | \n1.50 | \n287 | \n4.29 | \n
50 | \nCML 197 | \n3.39 | \nS | \n98.53 | \n19.67 | \n2.67 | \n213 | \n4.41 | \n
\n | LSD | \n0.396 | \n— | \n19.159 | \n9.013 | \n0.902 | \n47 | \n1.3 | \n
\n | Pr > f | \n** | \n— | \n** | \n** | \n* | \n** | \n** | \n
\n | CV (%) | \n10.1 | \n— | \n18.8 | \n32.1 | \n36.9 | \n11.9 | \n24.7 | \n
\n | Overall mean | \n2.43 | \n— | \n62.93 | \n17.31 | \n1.51 | \n245 | \n3.23 | \n
Maize leaf blight reactions, grain yield, and thousand seed weight of 50 inbred lines tested during 2012 at Bako research Center in Ethiopia.
DSS = disease severity score (0.00–5.00); R = resistant (1.0–2.00); I = intermediate (2.10–2.50); susceptible (2.51–5.00); and TSW = thousand seed weight.
*; ** = Significant at 0.05 and 0.01 probability levels, respectively.
The significant differences detected among genotypes in this study across the 2 years (cropping seasons) was attributable to a range of factors such as favorable climatic conditions, the inoculation method employed, and proper disease rating. In other studies, the development of NLB was attributed to pathogenic fitness and environmental conditions [34]. In Ethiopia, the disease infection and epidemics in maize occur largely during the main production season particularly in the wet and humid areas. Therefore, breeding for resistance to the disease in such areas is critical.
\nDisease severity scores in both cropping seasons were significantly different (P < 0.01) (\nTables 3\n and \n4\n). During the two seasons, the lowest severity scores were observed for the inbred lines CML 202, 144-7-b, and 142-1-e. In contrast, relatively high severity scores were observed for CML 197, Kuleni-C1-101-1-1-1, and SC-22-430(63), suggesting that they were susceptible to the disease. The final severity score and AUDPC values provided sufficient estimation of the reaction of the inbred lines to E. turcicum. The inbred lines that were classified as resistant showed significantly lower AUDPC values than the susceptible ones (\nFigure 1\n). Furthermore, susceptible inbred lines tended to show a rapid increase in severity of the disease compared with the resistant lines culminating in higher severity scores toward maturity unlike the resistant ones. The severity of the disease was slightly higher in 2011 than 2012 (\nTables 3\n and \n4\n). This was likely due to the low rainfall that was received at flowering in 2012, which was not conducive for the development of the disease. Nonetheless, the environmental conditions were generally favorable for leaf blight development during the two testing seasons. Previous studies involving leaf blight showed that the dropper inoculation was efficient and minimized the chances of disease escape from evaluation [9]. In this study, the inoculation technique was easy to employ and reliable. There were clear differences between resistant and susceptible genotypes, and at the flowering stage, the later genotypes exhibited a moderate increase in diseased leaf tissue. In some cases, relatively less susceptible individual genotypes were identifiable. The selection of such less susceptible genotypes can result in the accumulations of minor genes that can elevate the level of field resistance [35–37].
\nArea under disease progress curve for resistant (red) and susceptible (green) maize inbred lines inoculated with isolates of E. turcicum in the field.
The twenty SSR primers identified 108 alleles among the 20 maize inbred lines. Between 1 to 11 alleles were scored across the SSR loci (\nTable 5\n). Two loci (Phi 037, Umc1296) each revealed only a single allele. The maximum number of alleles (11) was detected at the Bnlg 2190 locus. The maximum PIC estimated for all loci was 0.8028 with a mean of 0.54 (\nTable 5\n). The expected heterozygosity (He) values, as a measure of allelic diversity at a locus, varied from 0.0000 to 0.8395 with an average of 0.5774. These values were well correlated with the number of alleles. Ten SSR loci (Umc1568, Nc003, Umc2214, Umc2038, Phi085, Umc1153, Bnlg238, Phi054, Bnlg2190, and Bnlg240) attained a PIC value >0.6, which indicated their potential to detect differences between the inbred lines.
\nSSR locus | \nRepeat types | \nBin number | \nNumber alleles | \nPIC value | \nHe | \n
---|---|---|---|---|---|
Umc1568 | \nTCG | \n1.02 | \n6 | \n0.6833 | \n0.7250 | \n
Bnlg176 | \n___ | \n1.03 | \n4 | \n0.3092 | \n0.3378 | \n
Bnlg182 | \n___ | \n1.03 | \n6 | \n0.5510 | \n0.5888 | \n
Phi 037 | \nAG | \n1.08 | \n1 | \n0.0000 | \n0.0000 | \n
Bnlg 108 | \n___ | \n2.04 | \n4 | \n0.4253 | \n0.4637 | \n
Nc003 | \nAG | \n2.06 | \n6 | \n0.7429 | \n0.7778 | \n
Umc2214 | \nCTT | \n2.1 | \n8 | \n0.7075 | \n0.7350 | \n
Bnlg602 | \n___ | \n3.04 | \n6 | \n0.4701 | \n0.4900 | \n
Umc2038 | \nGAC | \n4.06 | \n4 | \n0.6311 | \n0.6925 | \n
Phi085 | \nAACGC | \n5.06 | \n4 | \n0.6695 | \n0.7222 | \n
Umc1153 | \nTCA | \n5.09 | \n8 | \n0.6683 | \n0.7036 | \n
Bnlg238 | \n___ | \n6 | \n8 | \n0.7689 | \n0.7922 | \n
Umc1296 | \nGGT | \n6.07 | \n1 | \n0.0000 | \n0.0000 | \n
PhiI015 | \nAAAC | \n8.08 | \n7 | \n0.5112 | \n0.5938 | \n
Umc1367 | \nCTG | \n9.05 | \n2 | \n0.4949 | \n0.5850 | \n
Phi054 | \nAG | \n10.03 | \n6 | \n0.8028 | \n0.8255 | \n
Umc1677 | \nGGC | \n10.05 | \n7 | \n0.3047 | \n0.3750 | \n
Bnlg2190 | \nAG | \n10.06 | \n11 | \n0.8224 | \n0.8395 | \n
Bnlg240 | \n\n | 8.06 | \n7 | \n0.7777 | \n0.8025 | \n
umc2361 | \nCCT | \n8.06 | \n2 | \n0.3743 | \n0.4986 | \n
Information about the 20 SSR loci used in this study.
PIC = polymorphic information content and He = heterozygosity.
The genetic diversity of the germplasm is one of the most important factors limiting the number of alleles identified per microsatellite locus during screening. However, other factors such as the number of SSR loci and repeat types as well as the methodologies employed for the detection of polymorphic markers have been reported to influence allelic differences. In this work, the mean number of alleles (5.4) was in agreement with those reported in maize [38]. Similarly, values of number of SSR loci used in this study closely agreed with the findings reported previously [13, 39]. In addition, the mean PIC value determined in the present investigation was in agreement with the findings that were obtained in earlier studies that involving the use of SSR markers on maize inbred lines [40, 41]. The PIC value demonstrates the usefulness of the SSR loci and their potential to detect differences among the inbred lines based on their genetic relationships. The dinucleotide SSR loci (phi054, nc003, bnlg2190) identified the largest mean number of alleles (7.67) and mean PIC (0.79), as compared to tri-, tetra-, and penta-nucleotide repeats in the study, which was in close agreement with previous observations in maize [40, 42].
\nIn this study, automated analysis was used for screening the microsatellites, resolving allelic variation better than using gel electrophoretic analysis for instance. This may be particularly important for SSR loci containing dinucleotide repeats whose amplification products are between 130 and 200 bp, because PCR products differing by two base pairs cannot be resolved with agarose gel electrophoresis [40, 43].
\nThe ability to measure genetic distances between the inbred lines that reflect pedigree relationship ensures a more stringent evaluation of the adequacy of marker profile data; hence, the minimum genetic distance which was revealed between CML-202 and I100E-1-9-1-1-1-1-1 (0.28) was a good indication, confirming the power of SSR markers to distinguish closely related inbred lines. Similar findings were reported for maize inbred lines using SSR markers [44–46].
\nThe dendrogram obtained using the UPGMA clustering algorithm based on SSR data matrices grouped the inbred lines into five categories (\nFigure 2\n). This information, in combination with the pedigree records and combining ability tests, will be valuable for selecting (or identifying) optimal crosses and assigning inbred lines into heterotic groups. The greatest distance was found between the cluster containing the inbred line CML-202 line and the cluster of the inbred line Gibe-1-91-1-1-1-1. Cluster I consisted of inbred lines that are adapted to mid-altitude as well as some originating from CIMMYT. Most of the mid-altitude inbred lines in this group originated from the heterotic group Kitale Synthetic II and constitute the largest group in the cluster. In Cluster II, CIMMYT inbred lines CML312 and CML395 were grouped along with two local inbred lines, with two subdivisions in the main group. Cluster III contained two major subgroups, one containing CIMMYT inbred lines and the other containing local inbred lines. In terms of pedigree, these inbred lines are closely related and belong to the heterotic group AB, thus supporting the observation of a positive relationship between the pedigree and the SSR marker groupings in this study. In another cluster, two CIMMYT inbred lines (CML-443 and CML-197) were grouped closely, as revealed on the UPGMA dendrogram (\nFigure 2\n). These two inbred lines were also grouped in the same heterotic groups A and AB, based on their heterosis. Cluster V consisted of one CIMMYT inbred line and two locally adapted mid-altitude inbred lines. The separation of these elite mid-altitude maize inbred lines into genetically distinct groups may be associated with high heterotic response and increased combining ability useful for hybrid development.
\nDendrogram showing genetic relationship among 20 maize inbred lines tested using 20 SSR markers. The five clusters among the inbred lines are denoted from I to V.
The majority of the inbred lines (60%) that were evaluated in this study were previously developed by the national maize breeding program in Ethiopia. Because of the potential of encountering genetic admixtures or incomplete pedigree records in breeding programs, discrepancies in classification of germplasm may occur when comparing molecular results with classification based on pedigree relatedness. The effects of selection, genetic drift, and mutation may contribute to these discrepancies. The technique of clustering inbred lines can create apparent discrepancies, when one inbred line that is related to two inbred lines from separate clusters is then grouped with the inbred to which it is more closely related [40, 47]. Nonetheless, the SSR markers separated most of the inbred lines into distinguishable clusters, which generally agreed with the existing pedigree records and the findings that were reported previously [27, 42].
\nThe inbred lines showed significant differences in reaction to the leaf blight disease and were classified into three categories namely resistant, intermediate, or susceptible. The mean disease severity and upper leaf area infection varied from 2.04 to 3.25 and 3.3% to 100% respectively. Seven inbred lines were identified as potential sources of resistance to leaf blight for the genetic improvement of maize under the mid-altitude agroecology in Ethiopia. The genotyping detected 108 alleles and grouped the inbred lines into five clusters consistent with their pedigrees. The genetic grouping present in the population as determined in this study will be useful in the exploitation of tropical germplasm for hybrid maize breeding programs. The inbred lines that were identified as resistant to leaf blight can be considered as source material for disease resistance under the mid-altitude agroecological conditions in Ethiopia. The genetic grouping of the inbred lines was valuable information for future maize breeding programs. The use of SSR markers was able to provide complimentary information regarding the relatedness of the elite inbred lines that were evaluated. The high PIC value across all loci was strong evidence confirming the potential for SSR markers to discriminate between inbred lines of diverse sources and even between closely related genotypes. A number of loci that were identified with high PIC values indicated their usefulness for diversity analysis of maize inbred lines. The approach used in the study enables clear differentiation between inbred lines and their classification into distinct groups based on genetic distance estimates generated through selected polymorphic SSR primers.
\nThere will be merit in establishing resistance breeding program aimed at developing varieties with increased adult plant resistance to TLB in Ethiopia. Such varieties offer one of the most effective and affordable ways to overcome the problem of leaf diseases of maize in the mid-altitude agroecology in Ethiopia and similar environments in SSA. Therefore, further testing of the resistant germplasm identified in this study across more locations and seasons will also be merited.
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