Developmental patterns in the leaves of water stressed and unstressed soybean plants measured immediately after the termination of the water deficit stress experiment.
Water stress is one of the most important physiological stress factors that adversely affect soybeans in many critical aspects of their growth and metabolism. Soybean’s growth, development and productivity are severely diminished, when soil or cell water potential becomes inadequate to sustain metabolic functioning. However, little has been done to gather comprehensive information regarding the specific changes that occur in water-stressed plants at the anatomical and morphological level. In this study, deviations in root growth, shoot growth, stomatal conductance, yield components and anatomical features are reported. Treatments with two levels of water stress imposed by reducing irrigation (once in 7 days or once in 15 days) revealed that, all cultivars (Dundee, LS 677, LS 678, TGx 1740-2F, TGx 1835-10E and Peking) were highly susceptible to prolonged water stress, exhibiting severe dehydration and death. A 15.0 and 30.0% survival frequency was obtained in plants irrigated once in 7 days; LS 677 and Peking, respectively. Unlike many other stresses, water deficit did not only affect the density of stomata, but, photosynthesis was affected by the lower levels of tissue CO2. These results suggest that, balanced biochemical, physiological, anatomical and morphological regulations are necessary for increased growth and yields in soybean.
- water stress
Water stress is one of the most important constraints in the growth and development of plants. Water deficit stress, in particular, is a major problem in agriculture and most crop plants show high sensitivity to this kind of stress than any kind of abiotic constraint conditions. Crop plant growth and yields are severely impacted by inadequate supply of water, which result in decreased carbon assimilates contents. In addition, plants exposed to prolonged shortage in ground, surface or atmospheric water, known as drought are highly susceptible to pests and diseases. Mattson and Haack  provided evidence on the occurrence of fungi and insect induced stalk rots, wilts and foliar diseases in plants caused by drought stress. The prevalence in disease outbreak occurred in water-stressed plants compared to the normal water stress-free plants. Estimations of yield losses in soybeans compiled by Wrather and Koenning in the United States from 1996 until 2007 indicated that, the role of pathogens such as soybean cyst nematode, phytophthora root and stem rot, as well as charcoal rot that affected seedling development was exacerbated by the physical environmental stress conditions . Drought is, and continues to be an insidious hazard to plants, animals and human lives. Drought conditions in many regions worldwide are worsening due to various factors, some of which are caused by climate change. The increase in atmospheric CO2 level, currently estimated at about 380–400 ppm, and alterations in hydrological cycles make drought a recurring natural hazard world-wide [3, 4]. In this regard, plants undergo permanent or temporary damage to their morphological architecture, and their anatomical and physiological processes when exposed to dry and hot conditions. According to Shao et al. , water stress effects can be extended in plants to alter gene expression, change cellular metabolism, cause reduction in mitotic cell division activities in mesophyll tissues and other organs, as well as to cause the decrease in stomatal conductance . Scientific research showed that; drought stress causes imbalances in the natural status of the environment and drastically disrupts crop cultivation thus, threatening food security [7, 8]. Many regions have experienced the detrimental and severe effects of drought, particularly, populations in the developing countries. In the Southern African Development Community (SADC) region; poor rainfall conditions were recorded for the 2016/2017 agricultural season as a result of El Niño induced drought . FAO’s global information and early warning system in 2015 reported significant drought dating back to 1984 . The area data covered regions such as the United States, Semi-Arido of Brazil, Eastern Europe and African countries where, severe drought causing food crisis across Ethiopia, Kenya and Somalia resulted into the deaths of over 1 million people. Therefore, the continuing drop into below-normal annual rainfalls and increasing temperatures create the relevance to study and understand the morphological/anatomical changes that plants undergo to cope with environmental stresses. In cultivated crops such as soybean (
2. Analyses of soybean responses to water deficit stress
Plants experience water deficit stress when the amount of water in the cells and surrounding becomes limiting to growth and development. To investigate these effects, a study was conducted to primarily assess the influence of water stress on the growth of soybean; morphologically and anatomically, under greenhouse conditions. According to Lisar et al.  water deficit is caused by prolonged water shortage. In order to examine this stress, reduction in the frequency of irrigation was performed by limiting watering to once a week (WT 1) and once in 15 days (WT 2). Plants used for the control were watered daily, depending on soil moisture content in the plastic pots. For the growth of soybean plants, plastic containers filled with a mixture of 4:1 (v/v) fertile sandy-loam soil with vermiculite was used. Seeds of soybean cultivar Dundee, LS 678, LS 677, TGx 1740-2F, TGx 1835-10E and Peking were inoculated into the pots for germination and seedling emergence. At least one soybean plant was grown per pot with 20 replicates per cultivar, and allowed to grow up to V3 stage before imposing water stress. The morphological and physiological data were then recorded, which included plant height, number of leaves plant-1, number of braches plant-1, yield and yield components, average leaf area, root length and the micro-morphological characteristics of the stomata and trichomes were evaluated. Assessment of these characteristics was guided by the methods according to Cornelissen et al.  with modifications. To study stomatal and trichomes characteristics the microscopic slides were prepared by a protocol modified from Yeung’s  guide to study plant structures. Leaves of soybean plants from both the control and water-stressed plants (WT 1 and WT 2) were collected a week before the experiment was terminated. The experiment was terminated when the plants reached reproductive stage 4 (R4) of fruiting, involving maturity and seed filling. The free hand sectioning method by Yeung was used to study structural organisation of the root and stems, with section staining done using Toluidine blue O stain. Quantifying the chlorophyll content and leaf area is an important measurement for comparing plant growth, treated with different growth conditions. For leaf area assessment, leaf samples were randomly detached from the different cultivars, and their leaf area estimated as described by Richter et al. . Leaves were randomly sampled for estimation of chlorophyll content using a CCM 200 plus Chlorophyll Meter, Opti-Sciences.
3. Description of soybean morphology and anatomy
Plants are responsible for a number of essential ecological services. Plants are the main primary source of foods for humans and animals, supply oxygen, timber, medicine and also have ornamental value. The multiple and complex processes involving genetic, morphological, anatomical, physiological and biochemical mechanisms are responsible for the goods and services that plants provide. These functions are made possible by the architecture of the plant’s internal and external structures. Soybeans like other legumes and non-leguminous plants display different types of internal and external growth forms that functions together to provide these services. The external form include indeterminate, determinate and semi-determinate morphological growth habits, which typically take place in both the early and late maturity groups of varieties grown for commercial and subsistence farming . Soybean plants with determinate growth terminate their vegetative growth stage during the onset of the reproductive stage. In contrast, indeterminate varieties continue growing even during flower setting and anthesis. Anthesis is the period in which flowers developed during the reproductive stage of the plant’s life cycle begin to open. According to the NDSU  the semi-determinate growth habit lies between the polarity and growth of the other two growth habits (determinate and indeterminate form). The vegetative parts of soybean include the stem, leaves and the soil submerged roots. A few types of leaves can be found in soybean. The plant has trifoliate leaves, which are photosynthetic foliage with three leaflets. They have protective scale leaves which covers and protect young immature flowers before anthesis.
These scales are small bracts which appear subtending the yellow or purple flowers of soybeans . The special leaf types constituting the floral parts or inflorescence (raceme) can also be found. The vegetative stage is furthermore characterised by erect elongated stems, axillary buds, some viewed immediately above the cotyledons at the axil, unifoliate buds and the terminal buds (Figure 1a–c). Both young and old stems of soybeans are heavily covered by the epidermal hairs (trichomes) (Figure 1d). Even though soybean plants produce primary roots, originating from the seedling’s embryo; the roots have many branching secondary roots that slightly resemble fibrous root system in monocots. Most of the lateral roots are concentrated at the upper part of the root zone. As in most of the dicotyledonous plants, soybean’s body is made up of the three main tissue systems: dermal, ground and vascular (Figure 1e, f). The epidermis as the dermal tissue is the outermost single layer of cells derived from the protoderm, and in soybean it covers the plant for its entire life cycle. The three main types of epidermal cells found in soybean include trichomes and microscopic guard cells as well as the subsidiary cells of the stomata (Figure 2b–d). This layer of elongated and compactly arranged cells functions to protect soybean against water loss and harsh external environmental factors, including pathogens. Trichomes are unicellular or multicellular hairs occurring on shoot system of plants. On the roots, hairs are called root hairs. In leaves, this layer of cells is followed by the palisade parenchyma and spongy mesophylls.
The palisade and spongy cells are specialised tissues used by all eudicot plants with C3 pathway for photosynthesis and gaseous exchange in leaves, respectively . Soybean is one of the C3 plants which undergo photosynthetic carbon reduction and do not have a CO2 concentrating mechanism. It differs with grain crops such as maize, rice, sorghum and wheat C4 plants that concentrate CO2 by not salvaging carbon lost during photorespiratory carbon oxidation (PCO) cycle . But, the palisade and spongy tissues of soybean form the mesophyll, a ground tissue system of a leaf, which plays a critical role in carboxylation, reduction and regeneration processes during photosynthesis. In roots and stems, the fundamental (ground) tissue consists of non-protective and non-conductive simple cells of parenchyma, collenchyma and sclerenchyma (Figure 2e and f). Evert and Eichhorn  referred to this tissue system as the one most dominated by parenchyma cells, which are by far considered the most common ground tissue of the pith and cortex in roots and stems of soybean and other eudicots, as well as in the monocots. The vascular system is made up of conducting strands of phloem and xylem. These are principal water and food conducting tissue in all vascular seedless and seed plants.
3.1. Morphological changes due to water deficit stress
The morphological evidence gathered in this study has shown that soybean growth is highly sensitive to water deficit stress. All plants exposed to water deficit presented significant changes in their shoot and root morphology. Complete reduction in the number of new branches per plant, initiation of leaves and expansion of the lamina (measured by estimated leaf area) and the number of trifoliate leaves per plant was observed. Decreases in the assessed morphological characteristics were more predominant in plants subjected to stress for longer periods (WT 2) than those watered once a week (WT 1). Soybean cultivar Dundee, TGx 1740-2F, TGx 1835-10E and Peking produced significantly similar mean number of trifoliate leaves (about 4.0–5.0) in WT 2, when compared to about 5.0–6.0 trifoliate leaves obtained in WT 1 (Table 1). Leaf rolling and flipping were observed in some of the older leaves as a result of induced water stress. The negative effects of water stress on new leaf and branch formation was also reported by Mabulwana . Jaleel et al.  similarly added that, water stress decreases the elongation and expansion of stems and leaves. In contrast to observations made in all water stressed plants, the control exhibited normal shoot growth and the highest number of trifoliate leaves (Table 1).
|Soybean genotypes||Treatment plants 1||Treatment plants 2||Control plants|
|Mean no. of fully developed trifoliate leaves||Average leaf area (cm2)||Stomatal density (no. of stomata/ cm2)||Mean no. of fully developed trifoliate leaves||Average leaf area (cm2)||Stomatal density (no. of stomata/cm2)||Mean no. of fully developed trifoliate leaves||Average leaf area (cm2)||Stomatal density (no. of stomata/ cm2)|
According to Nosalewicz and Lipiec  suppression on the growth and distribution of the roots by water stress could also lead to the reduction in shoot growth. As the vegetative shoot growths appeared diminished by induced stress, roots in water-stressed plants became more elongated and branched than in the control. Root phenotype in the control appeared shallow and less branched than in WT 1 and WT 2 plants. However, plants which had irrigation reduced to once in 15 days (WT 2) had deep root phenotype compared to plants irrigated once a week (WT 1). Insufficient water supply for WT 2 plants with deep root development, and moderately stressed plants (WT 1), both demonstrated clear morphological changes. All cultivars in WT 2 also exhibited severe nutrient deficiency symptoms (the entire leaf with chlorosis and marginal necrosis) and stem wilting. These symptoms were accompanied by adverse growth effects and survival frequency of 0% when the experiment was terminated (Table 2). Water deficit stress ultimately led to the severe damage to shoots of WT 2 plants, with no possible indication of recovery. In WT 1 plants, moderate to severe deficiency symptoms were observed. Soybean cultivar LS 677 and Peking showed some resistance with 15 and 30% survival rate (Table 2). A few plants in these two genotypes exhibited moderate stress effects among all the cultivars assessed. There were no differences in the lengths of root system and shoots observed in water stress resistant cultivars (LS 677 and Peking) in comparison with those severely affected (Dundee, LS 678, TGx 1740-2F and TGx 1835-10E) in both WT 1 and WT 2 plants.
|Soybean genotypes||Mean plant height (cm)||Mean no. of branches||Flowering plants (%)||Mean no. of pods produced||Survival frequency (%)|
|TP 1||TP 2||TP 1||TP 2||TP 1||TP 2||TP 1||TP 2||TP 1||TP 2|
Klamkowski and Treder  reported almost similar results in water stressed strawberry plants. In addition, there were no major differences, especially in root lengths that were observed between water stressed plants and the control. The report cited inhibition of growth by water stressed plants, involving decrease in root expansion as suggested by Boyer . This claim probably led to the observed root phenotype in water stressed strawberry plants. This is in contrast with finding in this study and most of the other suggestions made on root phenotypes during water stress. In general, root formation has been found to increase in length during water stress, with roots growing deep into the soil in search for moisture [17, 19, 23, 24]. This further development in the root system is an attempt by plants to increase the uptake of water in order to sustain growth as observed in this study.
3.2. Anatomical changes in response to water stress
The WT 1 and WT 2 plants demonstrated a different internal anatomy compared to the control plants. Stem cortex of water stressed plant were generally smaller compared to the cortex in stems of plants in the control (Figure 2). However, vascular tissue thickening and expansion was observed in both the roots and stems of water stressed and control plants. The development of the secondary tissues in water stressed plants, especially the deposition of secondary xylem cells (as viewed in Figure 2a, b), was interrupted by the gradual rapturing of the pith which resulted in the formation of pith canals. Pith canals are hollow centres, called central canals, which are usually formed in woody shrubs and trees. They are formed when the earliest vascular tissues, protoxylem, is destroyed by the formation of new metaxylem as the root or stem grows in diameter. In gymnosperms, these canals are instead used by the pine trees to store resin and they are more associated with the cortical tissue of the stems than the pith . In stems of plants such as seedless vascular plants (horsetails), these canals are naturally formed to reduce the weight of the stem thus, increasing stem strength and resistance to buckling . However, the formation of canals (breaking down of the soybean pith tissue) observed in roots and stems may have resulted from water stress. Furthermore, this may have possibly impacted negatively on the growth of plants, particularly when induced as a result of severe water stress, like in WT 2.
Even though the pith is poor in nutrients , the parenchyma cells can still function in storage of nutrients and water for the plant. Pallardy  suggested that, rapturing could also destroy the interconnectivity between the storage parenchyma of the pith with the cortex, disrupting short distance transport that occurs through the rays via secondary xylem. The variations in canal diameters between WT 1 and WT 2 (Figure 2a, b, including canal in the root-
The reduction occurred when growth is affected by death of tissues and slowing down of metabolism as a result of the stress. Alteration in plant metabolism affect cell division, thus cell elongation and expansion is negatively affected as evidenced in Figure 2d, e). The xylem cell portion in the roots of water stressed plants was reduced compared to xylem tissue diameter in the control. Yamaguchi and Sharp  indicated that, water stress induce changes in root growth and cell length distribution which may be directly related to growth inhibition in roots, especially at root elongation zones. Another example is by Schuppler et al.  who also indicated the reduction on mitotic activity of mesophyll tissues in wheat (
4. Effect of water deficit on plant metabolism
The synthesis and breakdown of metabolites to yield energy is required for the many activities that plants depend upon. But, when plants are exposed to drought stress conditions, physiological and metabolic changes occur. Immediate acclimatisation by the alteration of plant morphology is therefore required for plants to be adapted to the changing environments. Whether plants succeed to acclimatise or not, the subsequent phenotypic modifications observed in water stressed plants would be a function of the metabolic changes. In soybean, like other leguminous plants, decrease in the leaf area, number of individual leaves and the total number of branches per plant is normally observed [16, 24]. However, on the metabolic section, water stressed plants experiences a dramatic decrease in photosynthetic rates as a consequence of the modification in photosynthetic structures and chloroplastidic pigments. Chloroplastidic pigments involve all plant pigments such as chlorophylls and carotenoid pigments embedded in the thylakoid membranes of parenchyma mesophylls . These pigments are primary molecules responsible for making sure that light energy from the sun is captured and converted to chemical energy required for metabolism.
This is the main route in which energy used for synthesis of biological products enters our biosphere. Water stress adversely limits this process by inhibiting the functioning of structure serving as primary support for photosynthetic metabolism. According to Kwon and Woo  drought reduce photosynthesis by limiting stomatal operations. In line with this report, the soybean plants subjected to water stress (WT 1 and WT 2) kept their stomata closed to reduce transpiration, hence trying to preserve water. The stomatal micrograph in Figure 3 illustrates closed stomatal apertures (c, d) prepared from leaves collected during the day. The closure of stomata in turn reduces the concentration of CO2 required in the mesophyll for carboxylation process during the manufacturing of photosynthates. This phenomenon was also reported by Dekov et al. , Evert and Eichhorn , Lopez-Carbonell et al.  and Taiz et al. . Additionally, there were significant variations in stomatal density exhibited by the different genotypes.
Water stressed soybean cultivar TGx 1740-2F and LS 677 exhibited low density of stomata with an average of 154 and 106 in WT 1 and WT 2 respectively, among all the cultivars used (Table 1). Furthermore, the two TGx cultivars (TGx 1740-2F and TGx 1835-10E) did not show extensive variations in the stomata among all water stressed plants, including the control. The mean leaf areas of the water stressed plants were also significantly lower compared to the control. The decrease in the leaf area of the plants posed negative effects on the rate of photosynthesis by reducing the leaf surface area in which light is captured. Anatomically, water stress also had an effect on leaf mesophyll thickness which also had an impact on photosynthesis. Cramer and Browman  attributed this to the changes in the rate of cellular expansion, which was observed in the maize mesophyll tissues when cell division and differentiation appeared affected by drought stress. However, plants growing in soil grounds of very lower water potential possess poor cell formation and expansion. Schuppler et al.  also reported this when assessing the effects of water stress on rate of cell division or mitotic activity on wheat leaf tissues. The report indicated that generally, leaf tissue expansion rate is reduced to more than 50% when plants are subjected to drought stress. In terms of physiological response to water stress, the reduction in chlorophyll content index (CCI) in water stressed plants was recorded, and the decreases in chlorophyll contents varied according to imposed water stress treatments (Figure 4). Plant irrigated once in 15 days (WT 2) showed remarkable decrease in CCI (Figure 4d) than WT 1 plants (Figure 4b).
Control plants did not exhibit significant reduction in CCI nor variation in all cultivars’ CCI measurements even before when water treatments were imposed on water stressed plants (Figure 4a, b). But then, differences were not expected in the CCI estimates of control plants measured early during growth and later before termination of the experiment, since the plants were adequately watered. Therefore, as expected the chlorophyll degradation was not induced on control plants as a result of water stress. As the differences in the chlorophyll content and degradation were observed in water stressed plants, these findings were in line with Dhanda et al.  and Benjamin and Nielsen ‘s reports on the effects of drought on plant metabolism. As indicated on Section 2, to examine and confirm the degradation of chlorophyll and its subsequent effects on photosynthetic activity, starch analysis was performed. Leaves detached from randomly selected soybean plants were obtained and taken to the laboratory for starch analysis. The leaves were bleached in boiling 90% ethanol and incubated in dilute iodine (0.5 M) solution (2:1) for 3 minutes and then rinsed with distilled water. Rinsing is necessary to remove excess iodine solution on the leaves while a colour change occurs. The iodine stained leaves (Figure 5a–c) were then visualised under a ZIESS Discovery V12 stereo microscope mounted with an ICc5 Axio-Camera. The presence of high starch content was observed in the control (Figure 5c); whereby starch contents in WT 1 (Figure 5a) and WT 2 (Figure 5b) were very drastically reduced because of poor photosynthetic activity. Intense blue-black colour on the leaves of control plants indicate the presence of starch, generated from the photosynthesised carbohydrates. Only minor traces of starch were observed from WT 1 and WT 2 leaves due to water stress.
The formation of cell protuberance containing nitrogen-fixing Gram-negative bacteria in the roots of legumes plays an important role in improving plant growth characteristics, crop productivity and maintaining soil fertility. This establishment of lumps on roots of plants (known as nodulation) guarantees the supply of fixed atmospheric N2 for use in the synthesis of proteins, nucleic acids and other necessary nitrogen-containing compounds required for plant, animal and human growth and development. However, various reports have indicated that, water stress induces low frequencies of nodulation in many legumes, including soybean. Miao et al.  provided evidence that verifies sensitivity of soybean nodulating root cells and Rhizobium to water stress. In 2003, Ramos et al.  also indicated that, water stress affect nodulation in other legume species like
6. Impact of water deficit on flowering and fruiting
The soybean genotypes showed great differences in the percentage flowering, pod formation and other yield related components. Water stressed plants produced less than 2% yield, in the two soybean cultivars (LS 677 and Peking) that survived induced water stress. A few WT 1 plants subjected to water stress continued their growth until flowering and pod formation stages. However, flower and fruit pod abortions were simultaneously observed leading to 7.0 and 3.0 mean pod number observed in the few plants that had survived (Table 3). These numbers were not comparable with the yield component data recorded for these cultivars in the control. Soriano et al.  determined a positive relationship between yield quantity by estimating grain number and weight in early planted sunflower by timing induction of environment stress. In line with this report, positive yield characteristics that include; total percentage of flowering plants, mean number of pods and average pod length, pod weight and seed weight (per 100 seeds) were observed in all of the cultivars in the control. In contrast, as a result of water stress, a significant number of flower abortions (10–15%) were observed in cultivar Dundee, LS 677, TGx 1740-2F and TGx 1835-10E which showed the least survival rate at 0%.
|Soybean genotypes||Mean plant height (cm)||Mean no. of branches||No. of flowering plants (%)||Mean no. of pods produced||Survival frequency (%)|
|Additional data on yield and yield components of untreated soybean plants|
|Ave. pod length (cm)||Ave. pod weight (g)||Seed weight/100 seeds (g)|
The variation observed in control plants however, did not seem to affect pod development and maturation, thus, could be attributed to the genotype performance than the environmental growth conditions (Table 3). In the control, a single genotypic setback was observed in cultivar TGx 1740-2F and TGx 1835-10E, which were the only ones producing the lowest number of pods, respectively. The effect of water stress in other oilseed grains such as sunflower, common bean, wheat, barley and maize were reported [41, 42, 43]. According to Jaleel et al.  the changes in the photosynthetic pigments and the decrease in metabolic functioning of the plant lead to decreased yield productivity. Seed yield and seed’s morphological characters can also be affected by drought . In cultivar Peking, the interaction between water deficit stress and seed appearance resulting from the genotype was not severely pronounced. The seeds appeared intensely shrinked and decreased in seed size due to loss of seed moisture, immediately after harvesting. This response was observed in another study assessing seed longevity in soybean seeds (data not published), clearly suggesting this as a dormancy or viability mechanism compared to other genotypes. In general, significant differences were observed during flowering, pod formation and seed maturation/ filling, as well as in the seed phenotypic characteristics among all cultivars in the control. Many water stressed plants (WT 1 and WT 2) did not survive to reach flowering as observed in the normally irrigated plants of the control (Tables 2 and 3).
7. Other biotic and abiotic stress effects
Plants are normally adapted to grow in complex and diverse environments. The success in growth establishment, reproduction and productivity of plant species rely upon a set of environmental conditions, natural resources and the interactions (beneficial or harmful) that exist among plants and other organisms. However, certain types of interactions, especially those including biological factors such as insects, parasites, viruses and bacterial pathogens have detrimental effects on plants. In addition, the non-living physical or chemical factors such as light, temperature, salinity, water, nutrient and other variables that can be found in the aquatic or terrestrial ecosystem also have major impacts on plant life. All above-mentioned factors may induce plant stress, defined by Taiz et al.  as a condition that prevent a given plant from achieving its maximum growth and reproductive potential as measured by vegetative growth, flowering, seed formation and yield quantity. Gerhardson  gave further information by providing more insights on disease symptoms caused by pathogenic strains of
Oilseeds such as soybeans have suffered major losses from the short and prolonged occurrence of abiotic stress, especially drought, extreme temperatures, flooding and waterlogging . Plants experiencing drought stress may also endure other stress effects simultaneously, like salinity and heat stress. Multiple stress effects and symptoms may be concomitantly induced by the occurrence of a single stress as described by Miransari  leading to combinational abiotic stress. In soybean, drought stress has many negative consequences ranging from reduced production of signalling and communication metabolites, decreased photosynthetic assimilates, nutrient deficiency, accumulation of reactive oxygen species (ROS) and reduction in nitrogen (N) fixation by affecting symbiotic relationship with
8. Water stress management and crop improvement
As previously discussed, drought stress is the most widely known and devastating stress factor that limit plant growth, development and productivity. Khaine and Woo  reported that, frequent drought effects recently and currently experienced, are largely induced by the changes in climatic conditions. The continuously fluctuating meteorological conditions in many regions worldwide have led to tremendous adversities on agriculture, biodiversity, wildlife and subsequently, the well-beings of many people. Plants normally evolve in order to adapt and adjust to the low water conditions or any other biotic and abiotic constraint. These adaptive measures are an important event of evolution in the history of life, with far reaching consequences as described by Kenrick and Crane . However, this is a very slow process in nature, even if it may result in greater diversity of plants, making changes in plants at their physiological, biochemical and molecular levels. These changes show a wide range of adaptations, at different levels in which plants attempt to deal with drought stress . Plants manage water stress in various ways. They regulate stomatal closure to reduce water loss, especially through transpiration. The stomatal opening and closing is very essential for gaseous exchange as reported by Osakabe et al. . They are controlled by complex regulatory events mediated by abscisic acid (ABA) signalling and ion transport induced by abiotic stress. Nonetheless, stomata closure negatively affects the rates of photosynthetic metabolism by lowering the amount of CO2. Plants also alter metabolic functions in order to inhibit the production of reactive oxygen species (ROS) such as superoxide (O2¯), and H2O2 [52, 58]. Other changes involve development of strategies to fix CO2 with minimum loss of water. For example; many C3 plants do not have photosynthetic adaptations to reduce the loss of CO2 molecules by separating photorespiration from the Calvin cycle. However, some succulent plants use Crassulaceae acid metabolism (CAM) to salvage CO2 minimising photorespiration thus, saving water.
In monocots such as maize and wheat, CO2 is fixed in the mesophyll spongy cells (a light-dependent process) and in the specialised cell around the leaf veins called the bundle sheath (light-independent). These monocotyledonous plants are referred to as C4 plants and they produce oxaloacetate which is converted into malate, transported into the bundle sheath for use in Calvin cycle [17, 59, 60]. Both C4 and CAM plants are well adapted to hot, dry environments than the C3 plants like peanut (
9. Conclusions and perspectives
This study revealed that soybeans are primarily affected by water deficit. Cultivars highly susceptible to water stress were easily distinguishable from those showing mild stress effects on the basis of the morphological and anatomical characters in stems, leaves and roots. Morphological architecture, anatomical features and chloroplastidic pigments were significantly affected by the induced water stress. This comprehensive insights regarding the internal and external growth characteristics including, aspects that involve the physiological processes is crucial for the pursuit of genetically modified plants. Soybean remains one of the most important oilseeds that are commercially and subsistently cultivated worldwide. The crop contains higher amounts of proteins, oils, fibre and minerals required for poultry feedstocks and human consumption. As a result of this, the elite superior genotypes of this crop still need to be investigated in order to identify the cultivars that would serve as genetic resource for breeding or genetic engineering, ultimately providing cultivars to be used for agricultural purposes showing high tolerance to abiotic stress, especially drought.
This study was carried out with the support by the Department of Biodiversity, University of Limpopo. The author acknowledges Dr. P.W. Mokwala and Mr. F. Nndwambi for providing seeds of soybean. Many thanks to Mr. L. Ramoba, F. Nukeri, S. Thosago, F.M. Masiya, R. Mudau, V.L. Makwana, L.R. Mahuluhulu, L.M. Marotola, K.K. Boshomane, M. Lutombo and T. Maleka for their technical support.
Mattson JW, Haack RA. The role of drought in outbreak of plant eating insects. Bioscience. 1987; 37(2):110-118
Wrather A, Koenning S. Effects of diseases on soybean yields in the United States 1996 to 2007. Plant Management Network. 2009; 401(01):1-7
Liu S, Yan D, Wang J, Wang B, Wang G, Yang M. Drought mitigation ability index and application based on balance between water supply and demand. Water. 2015; 7:1792-1807
Petit JR, Jouzel J, Raynaud D, Barkov NI, Barnola JM, Basile I, Bender M, Chappellaz J, Davis J, Delayque G, Delmolte M, Kotlyakov VM, Legrand M, Lipenkov V, Lorius C, Pepin L, Ritz C, Saltzman E, Stievenard M. Climate and atmospheric history of the past 420,000 years from the Vostok ice Core Antarctica. Nature. 1999; 399:429-436
Shao HB, Chu LY, Jaleel CA, Zhao CX. Water-deficit stress-induced anatomical changes in higher plants. Comptes Rendus Biologies. 2008; 331:215-225
Schuppler U, He PH, John PCL, Munns R. Effect of water stress on cell division and cell-division-cycle 2-like cell-cycle kinase activity in wheat leaves. Plant Physiology. 1998; 177:667-678
Jewell MC, Campbell BC, Godwin ID. Transgenic plants for abiotic stress resistance. In: Kole C, Michler C, Abbott AG, Hall TC, editors. Transgenic Crop Plants: Volume 1; Principles and Development. Berlin Heidelberg: Springer-Verlag; 2010. pp. 76-132
Lisar SYS, Motafakkerazad R, Hossain MM, Rahman IMM. Water stress in plants: Causes, effects and responses. In: Rahman MM, Hasegawa H, editors. Water Stress. Rijeka, Croatia: InTech Open; 2012
Food, Agriculture and Natural Resources (FANR). 2017. Food security; early warning system. Directorate for the Southern African Development Community (SADC). Issue no. 3
Food and Agriculture Organization of the United Nations (FAO). 2015. Monitoring agriculture drought with remote sensing data. FAO Agriculture Stress Index System
Ishaq MN, Ehirim BO. Improving soybean productivity using biotechnology approach in Nigeria. World Journal of Agriculture Sciences. 2014; 2(2):13-18
Cornelissen JHC, Lavorel S, Garnier E, Diaz S, Buchmann N, Gurvisch DE, Reich PB, Steege H, Morgan HD, van der Heijden MGA, Pausa JG, Poorter H. A handbook of protocols for standardized and easy measurements of plant functional traits worldwide. Australian Journal of Botany. 2003; 51:335-380
Yeung EC. A Beginner’s Guide to the Study of Plant Structure. Calgary Alberta, Canada: Association for Biology Laboratory Education (ABLE); 1998. pp. 125-136
Richter GL, Junior AZ, Streck NA, Guedes JVC, Kraulich B, Da-Rocha TSM, Winck JEM, Cera JC. Estimating leaf area of modern soybean cultivars by a non-destructive method. Crop Production and Management. 2014; 74(4):476-425
North Dakota State University (NDSU). Soybean Growth and Management: Quick Guide. Fargo, North Dakota: NDSU Agriculture Extension Service; 2015
Mabulwana PT. Determination of drought stress tolerance among soybean varieties using morphological and physiological markers. Masters’ Dissertation. Sovenga, South Africa: University of Limpopo; 2013
Taiz L, Zeiger E, Moller IM, Murphy A. Plant Physiology and Development. 6th ed. USA: Sinauer Associates; 2015. p. 561
Evert RF, Eichhorn SE. Raven Biology of Plants. 8th ed. New York: W.H. Freeman and Company; 2013. pp. 541-547
Jaleel CA, Manivannan P, Wahid A, Farooq M, Al-Juburi HJ, Somasundararam R, Panneerselvam R. Drought stress in plants: A review on morphological characteristics and pigments composition. International Journal of Agriculture and Biology. 2009; 11:100-105
Nosalewicz A, Lipiec J. The effect of compacted soil layers on vertical root distribution and water uptake by wheat. Plant and Soil. 2013; 375:229-240
Klamkowski K, Treder W. Morphological and physiological responses of strawberry plants to water stress. Agriculturae Conspectus Scientificus. 2006; 71(4):159-165
Boyer JS. Leaf enlargement and metabolic rates in corn, soybean and sunflower at various leaf water potentials. Plant Physiology. 1970; 46:233-235
Fenta BA, Beebe SE, Kunert KJ, Burridge JD, Barlow KM, Lynch JP, Foyer CH. Field phenotyping of soybean roots for drought stress tolerance. Agronomy. 2014; 4:418-435
Kwon MY, Woo SY. Plant’s responses to drought and shade environments. African Journal of Biotechnology. 2016; 15(2):29-31
Rosner S, Hannrup B. Resin canal traits relevant for constitutive resistance of Norway spruce against bark beetles: Environmental and genetic variability. Forest Ecology and Management. 2004; 200:77-78
Kirkendall LR, Biedermann PHW, Jordal BH. Evolution and diversity of bark and ambrosia bettles. In: Vega FE, editor. Bark Beetles: Biology and Ecology of Native and Invasive Species. USA: Academic Press; 2015. pp. 85-156
Pallardy SG. Vegetative growth. In: Physiology of Woody Plants. 3rd ed. USA: Academic Press; 2008. pp. 39-86
Yamaguchi M, Sharp RE. Complexity and coordination of root growth at low water potential: Recent advances from transcriptomic and proteomic analyses. Plant, Cell & Environment. 2010; 33:590-603
Munns R, Sharp RE. Involvement of abscisic acid in controlling plant growth in soil of low water potential. Australian Journal of Plant Physiology. 1993; 20(5):425-437
Lopez-Carbonell M, Alegre L, Vanonckelen H. Effect of water stress on cellular ultrastructure and on concentration of endogenous abscisic acid and indole-3-acetic acid in Fatsia jabonicaleaves. Plant Growth Regulation. 1994; 14:29-35
Dekov I, Tsonev T, Yordanov I. Effect of water stress and high-temperature stress on the structure and activity of photosynthetic apparatus of Zea maysand Helianthus annuus. Photosynthetica. 2000; 38:361-366
Cramer GR, Browman DC. Kinetics of maize leaf elongation: I. Increased yield threshold limits short-term, steady-state elongation rates after exposure to salinity. Journal of Experimental Botany. 1991; 42:1417-1426
Dhanda SS, Sethi GS, Behl RK. Indices of drought tolerance in wheat genotypes at early stages of plant growth. Journal of Agricultural Crop Science. 2004; 190:6-12
Benjamin JG, Nielsen DC. Water deficit effects on root distribution of soybean, field pea and chickpea. Field Crops Research. 2006; 97:248-253
Miao S, Jin J, Shi H, Wang G. Effect of short-term drought and flooding on soybean nodulation and yield at key nodulation stages under pot culture. Journal of Food, Agriculture and Environment. 2012; 10(3):819-824
Ramos MLG, Persons R, Sprent JI, James EK. Effect of water stress on nitrogen fixation and nodule structure of common bean. Pesquisa Agropecuária Brasileira. 2003; 38(3):339-347
Shetta ND. Influence of drought stress on growth and nodulation of Acacia origena(Hunde) inoculated with indigenous rhizobium isolated from Saudi Arabia. American-Eurasian Journal of Agricultural and Environmental Sciences. 2015; 15(5):699-706
Dupont L, Alloing G, Pierre O, Elmsehli S, Hopkins J, Herouart D, Frendo P. The legume root nodule from symbiotic nitrogen fixation to senescence. In: Nayata T, editor. Senescence. Rijeka, Croatia: InTech Open; 2012. pp. 137-168
Serraj R, Sinclair TR, Allen LH. Soybean nodulation and N2 fixation response to drought under carbon dioxide enrichment. Plant, Cell and Environment. 1998; 21:491-500
Stajkovic O, Delic D, Josic D, Kuzmanovic D, Rasulic N, Knezevic-Vukcevic J. Improvement of common bean growth by co-inoculation with Rhizobiumand plant growth-promoting bacteria. Romanian Biotechnological Letters. 2011; 16(1):5919-5926
Soriano MA, Villalobos FJ, Feres E. Stress timing effects on sunflower harvest index. In: Villalobos FJ, Testi L, editors. European Society for Agronomy, VII ESA Congress 15-18 July, Spain Book of Proceedings, 2002:142-143
Edward D, Wright D. The effects of winter water-logging and summer drought on the growth and yield of winter wheat ( Triticum aestivumL.). European Journal of Agronomy. 2008; 28:234-244
Lipiec J, Doussan C, Nosalewicz A, Kondracka K. Effect of drought and heat stress on plant growth and yield: A review. International Agrophysics. 2013; 27:463-477
Sepanlo N, Talebi R, Rokhzadi A, Mohammadi H. Morphological and physiological behaviour in soybean ( Glycine max) genotypes to drought stress implemented at pre- and post- anthesis stages. Acta Biologica Szegediensis. 2014; 58(2):109-113
Gerhardson JL. Pathogenicity of clover root pathogens to pea, bean and lucerne. Journal of Plant Disease and Protection. 2002; 109(2):142-151
Suleiman MN. Occurrence of Pythium aphanidermatumon cowpea ( Vigna unguiculata(L.) Walp) in Nigeria. Journal of Applied Biosciences. 2010; 26:1659-1663
Latef AAHA, Jan S, Abd-Allah EF, Rashid B, John R, Ahmad P. Soybean under abiotic stress. In: Azooz MM, Ahmad P, editors. Plant-Environment Interaction: Responses and Approaches to Mitigate Stress. UK: John Wiley and Sons, Ltd; 2016
Miransari M. Enhancing soybean response to biotic and abiotic stresses. In: Miransari M, editor. Abiotic and Biotic Stresses in Soybean Production, Soybean Production. Vol 1. UK: Academic Press Inc.; 2016. pp. 149-172
Bhatnagar-Mathur P, Vadez V, Sharma KK. Transgenic approaches for abiotic stress tolerance in plants: Retrospect and prospects. Plant Cell Reports. 2008; 27:411-424
Condon AG, Richards RA, Rebetzke GL, Farguhar GD. Breeding for high water-use efficiency. Journal of Experimental Botany. 2004; 55:2447-2460
Mutava RN, Prince SJK, Syed NH, Song L, Valiyodun B, Chen W, Nguyen HT. Understanding abiotic stress tolerance mechanisms in soybean: A comparative evaluation of soybean response to drought and flooding stress. Plant Physiology and Biochemistry. 2015; 86:109-120
You J, Chan Z. ROS regulation during abiotic stress responses in crop plants. Frontiers in Plant Sciences. 2015; 6(1092):1-15
Mangena P. Oryzacystatin 1 based genetic transformation in soybean for drought tolerance. Masters’ Dissertation. Sovenga, South Africa: University of Limpopo; 2015
Khaine I, Woo SY. An overview of interrelationship between climate change and forests. Forest Science and Technology. 2015; 11(1):11-18
Kenrick P, Crane PR. The origin and early evolution of plants on land. Nature. 1997; 389:33-39
Xoconostle-Cazares B, Ramirez-Ortega FA, Flores-Elenes L, Ruiz-Medrano R. Drought tolerance in crop plants. American Journal of Plant Physiology. 2011; 5(5):1-15
Osakabe Y, Osakabe K, Shinozaki K, Tran LSP. Response of plants to water stress. Frontiers in Plant Science. 2014; 5(86):1-8
Gill SS, Tuteja N. Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology and Biochemistry. 2010; 48(12):909-930
Hopkins WG. Introduction to Plant Physiology. 2nd ed. USA: John Wiley and Sons, Inc; 1999. pp. 189-214
Simpson MG. Plant Systematics. USA: Elsevier Inc; 2010. pp. 535-539
Zhang JZ, Creelman RA, Zhu JK. From laboratory to field. Using information from Arabidopsisto engineer salt, cold and drought tolerance in crops. Plant Physiology. 2004; 135(2):615-621
Montero-Tavera V, Ruiz-Medrano R, Xoconostle-Cazares B. Systematic nature of drought-tolerance in common bean. Plant Signaling & Behavior. 2008; 3(9):663-666
Abenavoli MR, Leone M, Sunseri F, Bacchi M, Sorgona A. Root phenotyping for drought tolerance in bean landraces from Calabria (Italy). Journal of Agronomy and Crop Science. 2015; 202(1):1-12
Nakashima K, Suenaga K. Review: Toward the genetic improvement of drought tolerance in crops. Japan International Research Quarterly. 2017; 51(1):1-10
Liu Z, Jin-Park B, Kanno A, Kameya T. The novel use of combination of sonication and vacuum infiltration in Agrobacterium-mediated transformation of kidney bean ( Phaseolus vulgarisL.) with lea gene. Molecular Breeding. 2005; 16:189-197
Saijo Y, Hata S, Kyozula J, Shimamoto K, Izui K. Over-expression of a single Ca2+-dependent protein kinase confers both cold and salt/ drought tolerance on rice plants. The Plant Journal. 2000; 23(3):319-327
Shou H, Bardallo P, Wang K. Expression of the Nicotianaprotein kinase (NPK1) enhanced drought tolerance in transgenic maize. Journal of Experimental Botany. 2004; 55(399):1013-1019
Ku YS, Au-Yeung WK, Yung YL, Li MW, Wen CQ, Liu X, Lam HM. Drought stress and tolerance in soybean. In: Board JE, editor. A Comparative Survey of International Soybean Research: Genetics, Physiology, Agronomy and Nitrogen Relationships. Rijeka, Croatia: InTech Open; 2013. pp. 209-237