Screening techniques and sources of resistance to abiotic stresses in cool-season food legumes

Summary The adaptability and productivity of cool-season food legumes (chickpea, faba bean, lentil, pea) are limited by major abiotic stresses including drought, heat, frost, chilling, waterlogging, salinity and mineral toxicities. The severity of these stresses is unpredictable in field experiments, so field trials are increasingly supplemented with controlled-environment testing and physiological screening. For drought testing, irrigation is used in dry fields and rain-out shelters in damp ones. Carbon isotope discrimination (Δ¹³C) is a well-established screen for drought tolerance in C3 cereal crops which is now being validated for use in grain legumes, but it is relatively expensive per sample and more economical methods include stomatal conductance and canopy temperature. Chickpea lines ICC4958 and FLIP87-59C and faba bean line ILB938 have demonstrated good drought tolerance parameters in different experiments. For frost tolerance, an efficient controlled-environment procedure involves exposing hardened pot-grown plants to sub-zero temperatures. Faba beans Cote d’Or and BPL4628 as well as lentil ILL5865 have demonstrated good freezing tolerance in such tests. Chilling-tolerance tests are more commonly conducted in the field and lentil line ILL1878 as well as derivatives of interspecific crosses between chickpea and its wild relatives have repeatedly shown good results. The timing of chilling is particularly important as temperatures which are not lethal to the plant can greatly disrupt fertilization of flowers. Salinity response can be determined using hydroponic methods with a sand or gravel substrate and rapid, efficient scoring is based on leaf symptoms. Many lines of chickpea, faba bean and lentil have shown good salinity tolerance in a single article but none has become a benchmark. Waterlogging tolerance can be evaluated using paired hydroponic systems, one oxygenated and the other de-oxygenated. The development of lysigenous cavities or aerenchyma in roots, common in warm-season legumes, is reported in pea and lentil but is not well established in chickpea or faba bean. Many stresses are associated with oxidative damage leading to changes in chlorophyll fluorescence, membrane stability and peroxidase levels. An additional factor relevant to the legumes is the response of the symbiotic nitrogen-fixing bacteria to the stress.     

Screening techniques and sources of resistance to foliar diseases caused by major necrotrophic fungi in grain legumes

Summary Necrotrophic pathogens of the cool season food legumes (pea, lentil, chickpea, faba bean and lupin) cause wide spread disease and severe crop losses throughout the world. Environmental conditions play an important role in the development and spread of these diseases. Form of inoculum, inoculum concentration and physiological plant growth stage all affect the degree of infection and the amount of crop loss. Measures to control these diseases have relied on identification of resistant germplasm and development of resistant varieties through screening in the field and in controlled environments. Procedures for screening and scoring germplasm and breeding lines for resistance have lacked uniformity among the various programs worldwide. However, this review highlights the most consistent screening and scoring procedures that are simple to use and provide reliable results. Sources of resistance to the major necrotrophic fungi are summarized for each of the cool season food legumes. Marker-assisted selection is underway for Ascochyta blight of pea, lentil and chickpea, and Phomopsis blight of lupin. Other measures such as fungicidal control and cultural control are also reviewed. The emerging genomic information on the model legume, Medicago truncatula, which has various degrees of genetic synteny with the cool season food legumes, has promise for identification of closely linked markers for resistance genes and possibly for eventual map-based cloning of resistance genes. Durable resistance to the necrotrophic pathogens is a common goal of cool season food legume breeders.     

Phenotyping of faba beans (<Emphasis Type="Italic">Vicia faba</Emphasis> L.) under cold and heat stresses using chlorophyll fluorescence

Temperature stress including low and high temperature adversely affect the growth, development and productivity of crops. Faba bean (Vicia faba L.) is an important crop as both human food source and animal feed, which contains a range of varieties that are sensitive to cold and heat stresses. In this study, 127 faba bean genotypes were collected from gene banks based on differences in geographical origin. The 127 genotypes were treated by single cold stress (2/2 °C day/night temperature (DT/NT)) and 42 genotypes were treated by either single episode of cold or heat (38/30 °C DT/NT) stress, or a combination of both at photosynthetic photon flux density of 250 µmol m^?2 s^?1. Chlorophyll fluorescence was used to detect the tolerance of faba beans to low and high temperatures. The maximum quantum efficiency of photosystem II (PSII), F_v/F_m, revealed pronounced differences in cold tolerance among the faba bean genotypes. The 42 genotypes were clustered into four groups according to cold and heat stresses, respectively, and the susceptibilities of faba beans under temperature stress could be distinguished. The combination of cold and heat stresses could aggravate the damage on reproductive organs, but not on the leaves, as indicated by the F_v/F_m. These results confirm that the use of F_v/F_m is a useful approach for detecting low and high temperature damage to photosystem II and to identify tolerant faba bean genotypes, however the results also indicate that the geographical origin of the genotypes could not directly be used to predict climate resilience. These sources of cold- and heat-tolerance could improve the temperature tolerance of faba bean in breeding programs.     

Screening techniques and sources of resistance to foliar diseases caused by fungi and bacteria in cool season food legumes

Screening techniques are an important component of the overall strategy of breeding for resistance to diseases in cool season food legumes. Suitable screening methods have been developed for several major foliar diseases of chickpea, pea, faba bean, and lentil, and sources of resistance have been identified. International cooperation plays an important role in promoting research and keeping collections of cultivated species and their wild relatives. New biotechnological approaches are promising for enhancing the practical use of genes for resistance.     

Evaluation of wild Arachis species for abiotic stress tolerance: I. Thermal stress and leaf water relations

The Indian groundnut cultivars have a narrow genetic base. Hence, it was of interest to investigate the genetic variability among wild Arachis species and their accessions for tolerance to thermal stress. A wide variation was observed in leaf morphological characters such as colour, shape, hairiness, length and width and thickness (SLA). The temperature and time required for 50% leaf injury was worked out with limited number of genotypes and was found to be 54°C for 50 min. Among 36 genotypes (having SLA in the range of 66 and 161 cm² g⁻¹) screened, the inherent potential for cold as well as heat tolerance in terms of relative leaf injury (RI) was observed. Thus, based on RI-values, A. glabrata 11824 and A. paraguariensis 12042 were identified as heat-tolerant and cold-tolerant genotypes, respectively while A. appresipila 11786 was found to be susceptible to both heat and cold. Correlation between SLA and RI values for heat (r = 0.38, P < 0.05) and cold (r = 0.52, P < 0.05) tolerance was positive, indicating that thicker the leaf the lower the injury or higher the tolerance. Among six species and 13 accessions, comprising both heat-tolerant and heat- susceptible genotypes, the concentrations of various leaf chemical constituents such as total protein, phenols, sugars, reducing sugar, amino acids, proline, epicuticular wax load and chlorophyll varied significantly. The epicuticular wax load ranged between 1.1 and 2.5 mg dm² among 13 A.glabrata accessions. These accessions were categorized into two groups, i.e. high-wax (range: 2.0–2.5 mg dm²) and low-wax types (range: 1.1–1.6 mg dm²). The high-wax type showed a higher diffusion resistance (dr) as compared to low-wax type; though the transpiration rate (tr) in high-wax type was moderate (between 9.5 and 11.6 μg cm⁻² s⁻¹). Genetic variability in parameters such as canopy temperature, dr and tr was also distinct. The fully turgid leaves with relative water content ≥91%, showed leaf water potential (ψ_leaf) between −0.7 and −1.2 MPa. Results indicated that the plants with thicker leaves are better protected from heat injuries. Further, epicuticular wax load seems to help in maintaining stomatal regulation and leaf water relations, thus affording adaptation to wild Arachis species to thrive under water-limited environments. The sources of tolerance, as identified in this study, could be utilized to improve thermal tolerance of the groundnut cultivars by intra-specific hybridization, following either conventional breeding using embryo rescue techniques, if required or utilizing biotechnological tools.     

A cytoplasmic-nuclear male-sterility system derived from a cross between <Emphasis Type="Italic">Cajanus cajanifolius</Emphasis> and <Emphasis Type="Italic">Cajanus cajan</Emphasis>

Cytoplasmic-nuclear male-sterility is an important biological tool, which has been used by plant breeders to increase yields in cross-pollinated cereals and vegetables by commercial exploitation of the phenomenon of hybrid vigor. In legumes, no such example exists due to the absence of an economic way of mass pollen transfer from male to female parent. Pigeonpea [Cajanus cajan (L.) Millsp.], however, is a different legume where a moderate level of insect-aided natural out-crossing (25–70%) exists and it can be used to produce commercial hybrid cultivars, if an efficient and stable cytoplasmic-nuclear male-sterility (CMS) system is available. This paper reports the development of a stable CMS system (ICP 2039A), derived from an inter-specific hybrid of Cajanus cajanifolius, a wild relative of pigeonpea, with a cultivar ICP 11501. Using this genetic material, designated as the A₄ cytoplasm, a number of fertility restorers and maintainers have been developed. The best short-duration experimental pigeonpea hybrid ICPH 2470 produced 3205 kg ha⁻¹ grain yield in 125 days, exhibiting 77.5% advantage over the control cultivar UPAS 120. At present, all the important biological systems necessary for a successful commercial hybrid breeding program are available in pigeonpea and the package of this technology has been adopted by private seed sector in India for the production and marketing of hybrid varieties.     

Effects of Salt Stress on Some Physiological and Photosynthetic Parameters at Three Different Temperatures in Six Soya Bean (Glycine max L. Merr.) Cultivars

The effect of NaCl (?0.1, ?0.4 and ?0.7 MPa) on some physiological parameters in six 23‐day‐old soya bean cultivars (Glycine max L. Merr. namely A 3935, CX‐415, Mitchell, Nazl?can, SA 88 and Türksoy) at 25, 30 and 35 °C was investigated. Salt stress treatments caused a decline in the K⁺/Na⁺ ratio, plant height, fresh and dry biomass of the shoot and an increase in the relative leakage ratio and the contents of proline and Na⁺ at all temperatures. Effects of salt stress and temperature on Chl content, Chl a/b ratio (antenna size) and qN (heat dissipation in the antenna) varied greatly between cultivars and treatments; however, in all cases approximately the same qP value was observed. It indicates that the plants were able to maintain the balance between excitation pressure and electron transport activity. Pigment content and the quantum efficiency of photosystem II exhibited significant differences that depended on the cultivar, the salt concentration and temperature. The cultivars were relatively insensitive to salt stress at 30 °C however they were very sensitive both at 25 and 35 °C. Of the cultivars tested CX‐415 and SA 88 were the best performers at 25 °C compared with SA 88 and Türksoy at 35 °C.     

Chickpea and faba bean nitrogen fixation in a Mediterranean rainfed Vertisol: Effect of the tillage system

Efficient management of legumes in order to maximize benefits depends on a correct field assessment of N₂ fixation. A field experiment was conducted during a 6-year period (2001–2002 to 2006–2007) in Córdoba (Southern Spain) on a rainfed Vertisol within the wheat-chickpea and wheat-faba bean rotation framework of a long-term experiment started in 1986. The aim was to determine the effect of tillage systems [no tillage (NT) and conventional tillage (CT)] on chickpea and faba bean N₂ fixation. Fixation was calculated using the ¹⁵N isotopic dilution (ID) and ¹⁵N natural abundance (NA) methods with the reference being the wheat crop. The strong inter-annual rain variation caused great differences in the behaviour of both leguminous plants with regard to grain yield, nodule biomass and N₂ fixation. The NT system showed more nodule biomass than the CT system in both legumes. The ID method was more accurate than the NA method in determining N₂ fixation. The average amount of fixed N in faba bean (80 kg ha^?1 year^?1) was much greater than that in chickpea (31 kg ha^?1 year^?1). The Vertisol under the NT system offered more favourable conditions for the stimulation of the N₂ fixation, with fixed N values that were significantly higher than under CT. The N added to the system through N₂ fixation was low in faba bean and virtually nonexistent in chickpea, only in terms of above-ground biomass.     

Heat stress in food legumes: evaluation of membrane thermostability methodology and use of infra-red thermometry

Heat tolerance in 45 chickpea, lentil, and faba bean genotypes was investigated during 2007/2008 and 2008/2009 at Alexandria Agriculture Research Station, Alexandria, Egypt, using screening methods employing the membrane thermostability technique. Threshold temperature to be used in screening for heat tolerance at germination was also investigated for each crop. Temperatures, responsible for 50% germination were 40, 33.5, and 29°C for chickpea, faba bean, and lentil, respectively. Germination percent under high temperature varied significantly (P ≤ 0.05) amongst genotypes. Germination percentage ranged from 4.8 to 71.6, 39.2 to 90.0, and 4.8 to 68.6, in chickpea, lentil, and faba bean, respectively. Differences were significant (P ≤ 0.05) among faba bean and chickpea genotypes. Membrane relative injury (RI%) showed significant (P ≤ 0.05) variability among the genotypes and ranged from 10.57 to 58, 5.2 to 61.7, and 15.7 to 52.7 in chickpea, lentil, and faba bean, respectively. Canopy temperature was measured to evaluate heat avoidance in tested genotypes. Infra-red thermometry was used to measure canopy temperature and the gradient of canopy to ambient air temperature (∆T_C-A) in moisture stressed and unstressed treatments. Canopy temperature, leaf water potential (LWP) and leaf water content were affected by the level of soil moisture. Genotypes were able to bring their canopy temperatures to levels lower than ambient air temperatures but the differences were not significant. A heat stress index (HSI) were computed relating the ∆T_C-A in moisture stressed to unstressed treatments. Regression of leaf water potential (LWP) and the heat stress index (HSI) was significant (P ≤ 0.05) in faba bean genotypes in the stressful environment. The results of the present investigation emphasize the efficiency of membrane thermostability technique in selection for heat tolerance in early stages of growth in food legumes.     

The combined effect of plant stature and maturity on the response of wheat and triticale accessions to Septoria tritici

Summary The relationships between percent pycnidia coverage on the four uppermost leaves (PCD), plant height (PHT) and days to heading (HED) were evaluated for 21,000 wheat and triticale accessions tested in artificially inoculated (with fixed combination of S. tritici isolates) field nurseries over 8 trial years. A general Linear Model procedure (GLM) estimated Septoria severity using two correlative models: model 133-1 Year and model II–PCD=b₁PHT+b₂HED+C. The regression coefficients for PHT and HED in the two models were –0.54 and –0.40, respectively, with a R²=0.80^** and R²=0.29^** for model I and model II, respectively. The predicted cultivar best fitted to the model would be characterized as a semidwarf (PHT=115 cm) with an early-moderate maturity (HED=95 days to heading). The estimated mean percent pycnidial coverage for the two models over the 8 trial years was 40.8%. The performance of a group of 38 cultivars replicated yearly during the 8 trial years was assessed relative to model I. The deviation of each cultivar from the model was calculated using two functions: a) Sum Relative Serial Deviation (SRSD) and b) Total Relative deviation (TRD), in addition to Standard errors (SE). The proposed analytical protocol enabled identification of cultivars which expressed consistent yearly deviation (from the model) in host response combined with low-moderate mean pycnidial coverage (±30%). Such cultivars may possess a more stable type of genetic protection against the adverse effects of septoria tritici blotch.     

Mutations affecting nodulation in grain legumes and their potential in sustainable cropping systems

Many spontaneous and a large number of induced mutants that show altered nodulation pattern have been isolated in pea, soybean, common bean, faba bean, chickpea, groundnut and pigeonpea. Available information on nodulation mutants in these crops is summarised. The importance of nodulation mutants in basic studies on plant-microbe symbiotic interactions, nitrogen fixation and breeding of cultivars with higher yield and nitrogen fixation rate are examined. The nodulation mutants, after inoculation with specific bacterial strains or a number of different strains, show either: no nodulation (nod-), few nodules (nod+/-), ineffective nodulation (fix-), hyper nodulation (nod++) or hypernodulation even in the presence of otherwise inhibitory nitrate levels (nts). No spontaneous hypernodulation or nts mutants have been found, all have been induced in independent experiments using different cultivars of pea, soybean and common bean after mutagenising seeds. Most nodulation mutants show monogenic recessive inheritance, though semi-dominant and dominant inheritance is also reported. Nodule number is controlled by a process known as autoregulation; hypernodulating mutants show relaxed autoregulation. By grafting shoots of hypernodulating soybean mutant on normal nodulating soybean, mungbean and hyacinth bean, presence of a common, translocatable signal has been shown. Nodulation mutants have contributed to the understanding of the genetic regulation of host-symbiont interactions, nodule development and N fixation. Initially, the hypernodulating mutants were found to be poor in yield. Using the induced hypernodulating mutant, a new soybean cultivar ‘Nitrobean 60’, has been released in Australia. This cultivar is reported to have given 15% higher yield over cv. ‘Bragg,’ and contributed a higher amount of fixed N to the following cereal crop in rotation. Prospects of using the nodulation mutants in developing grain legume cultivars that combine high yield with high residual N, within the bioenergetic constraints, for developing sustainable cropping systems are examined. This revised version was published online in August 2006 with corrections to the Cover Date.     

Breeding high-protein pigeonpea genotypes and their agronomic and biological assessments

Proteins, inevitable for nutritional security of human beings and legumes, by far, are the cheapest source of this vital nutrient. The escalating prices and never halting population growth limit the per capita availability of protein-rich legumes. In view of limited land resource and need to grow other food crops, the greater protein harvests are possible only by increasing the protein levels of popularly grown legumes. In this context, attempts were made for raising the protein content in pigeonpea [Cajanus cajan (L.) Millsp.] through traditional plant breeding tools. For this, the high-protein trait was successfully transferred from wild relatives of pigeonpea to the cultivated types. In the derived inbred lines, the protein content was significantly enhanced from 20% - 22% to 28% - 30%. Two high-protein lines HPL 40 and HPL 8 also produced 2100 and 1660 kg/ha grain yield, respectively. This simply means that, in comparison with traditional cultivars, the cultivation of high-protein lines will provide additional 100 kg/ha of digestible protein to the farming family. This paper, besides describing the breeding procedures, also discusses the accomplishments of this breeding endeavour with respect to its various nutritional and biological properties.     

Differential heat sensitivity of two cool-season legumes,chickpea and lentil,at the reproductive stage,is associated with responses in pollen function,photosynthetic ability and oxidative damage

Increasing temperatures are adversely affecting various food crops, including legumes, and this issue requires attention. The growth of two cool-season food legumes, chickpea and lentil, is inhibited by high temperatures but their relative sensitivity to heat stress and the underlying reasons have not been investigated. Moreover, the high-temperature thresholds for these two legumes have not been well-characterised. In the present study, three chickpea (ICCVO7110, ICC5912 and ICCV92944) and two lentil (LL699 and LL931) genotypes, having nearly similar phenology with respect to flowering, were grown at 30/20°C (day/night; control) until the onset of flowering and subsequently exposed to varying high temperatures (35/25, 38/28, 40/30 and 42/32°C; day/night) in a controlled environment (growth chamber; 12 hr/12 hr; light intensity 750 µmol m⁻² s⁻¹; RH-70%) at 108 days after sowing for both the species. Phenology (podding, maturity) was accelerated in both the species; the days to podding declined more in lentil at 35/25 (2.8 days) and 38/28°C (11.3 days) than in chickpea (1.7 and 7.1 days, respectively). Heat stress decreased flowering–podding and podding–maturity intervals considerably in both the species. At higher temperatures, no podding was observed in lentil, while chickpea showed reduction of 14.9 and 16.1 days at 40/30 and 42/32°C, respectively. Maturity was accelerated on 15.3 and 12.5 days at 38/28°C, 33.6 and 34 days at 40/30°C and 45.6 and 47 days at 42/32°C, in chickpea and lentil, respectively. Consequently, biomass decreased considerably at 38/28°C in both the species to limit the yield-related traits. Lentil was significantly more sensitive to heat stress, with the damage—assessed as reduction in biomass, reproductive function-related traits (pollen viability, germination, pollen tube growth and stigma receptivity), leaf traits such as membrane injury, leaf water status, photochemical efficiency, chlorophyll concentration, carbon fixation and assimilation, and oxidative stress, appearing even at 35/25°C, compared with 38/28°C, in chickpea. The expression of enzymatic antioxidants such as superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase and non-enzymatic antioxidants declined remarkably with heat stress, more so in lentil than in chickpea. Carbon fixation (assessed as Rubisco activity) and assimilation (assessed as sucrose concentration, sucrose synthase activity) were also reduced more in lentil than in chickpea, at all the stressful temperatures, resulting in more inhibition of plant biomass (shoot + roots), damage to reproductive function and severe reduction in pods and seeds. At 38/28°C, lentil showed 43% reduction in biomass, while it declined by 17.2% in chickpea at the same time, over the control temperature (30/20°C). At this temperature, lentil showed 53% and 46% reduction in pods and seed yield, compared to 13.4% and 22% decrease in chickpea at the same temperature. At 40/30°C, lentil did not produce any pods, while chickpea was able to produce few pods at this temperature. This study identified that lentil is considerably more sensitive to heat stress than chickpea, as a result of more damage to leaves (photosynthetic ability; oxidative injury) and reproductive components (pollen function, etc.) at 35/25°C and above, at controlled conditions.     

Variation in Cicer arietinum L.

Summary A collection of populations and cultivars of Cicer arietinum L. were studied to obtain phenotypic, genotypic and environmental correlation coefficients, and broad sense heritabilities. Principal Component Analyses were performed on phenotypic, genotypic and environmental matrices. Data and phase obtained on a pure morphological basis, as well as on quantitative genetic studies and geographical distribution support the existence of two complexes within the cultivated chickpea, macrosperma and microsperma. These taxa differ in a cluster of complex characters associated with seed. pod and leaf morphology, and they differ in distribution. There is no taxonomic basis to treat these as subspecies. We propose to include C. reticulatum Lad., the wild chickpea, as a subspecies of C. arietinum, with the cultivated kinds recognized as subspecies arietinum. Race macrosperma was derived from race microsperma through selection during relatively recent times of the evolutionary history of the chickpea.     

灌浆期干旱胁迫对玉米叶片关系统活性的影响

以高淀粉玉米品种郑单21为材料,借助叶绿素荧光快速诱导动力学曲线和820 nm光吸收曲线,研究了灌浆期土壤干旱胁迫对玉米籽粒产量和对叶片光系统I (PSI)及光系统II (PS II)活性的影响。两年的研究结果均表明,干旱胁迫显著抑制叶片光合速率(P<0.05)和籽粒产量(P<0.05)。JIP-test分析发现干旱胁迫导致叶绿素荧光快速诱导动力学曲线中的K点和J点上升,表明PS II放氧复合体(OEC)和Q_A之后的电子传递链受到抑制,且PS II受体侧受抑制的程度大于供体侧。此外,干旱胁迫也显著地抑制PS I的最大氧化还原活性(ΔI/I_o),阻碍光合电子从PS II向PS I的传递,破坏了PS I和PS II的协调性。我们认为干旱胁迫抑制PS I和PS II活性并破坏二者的协调性,是导致导致P_n和籽粒产量下降的重要原因之一。     

Allozyme evaluation of upright common bean genotypes

Summary In common bean (Phaseolus vulgaris L.) the diaphorase (DIA) allozyme variant Diap-2 ¹⁰⁵ is frequently present in plants with upright, type II plant architecture. The genetic relationship between upright plant architecture and Diap-2 ¹⁰⁵ was investigated in eight F₂ populations derived from crosses between navy bean and pinto bean parental lines differing for type I, II, and III growth habit and DIA genotype. Segregation at the Diap-2 locus followed the expected 1:2:1 ratio in all eight F₂ populations and when pooled across F₂ populations. F₂ data from 1345 individuals indicated that plant architecture and the Diap-2 locus are not linked (r=0.03, P=0.333). However, the Diap-2 ¹⁰⁵ allozyme was present in 71% of advanced navy, pinto, and great northern genotypes with type II plant architecture. Due to random drift, Diap-2 ¹⁰⁵, initially associated with type II architecture through founder effect, may be maintained in such genotypes without providing greater fitness or without being associated with a locus or linked loci governing upright plant architecture.     

Conservation of microsatellite flanking sequences in different taxa of Leguminosae

To test if locus-specific microsatellite markers designed for one genus are informative when used with related genera, the conservation of microsatellite-flanking intergeneric primer binding sites was tested in the closely related tribes Vicieae and Cicereae, from the subfamily Papilionoideae of the Leguminosae family. A total of 123 sequence-tagged microsatellite sites (STMS) markers derived from chickpea were used to amplify loci in lentil (Lens) and dry pea (Pisum). The percentage of chickpea primer binding sites conserved between the three genera was 54.4%. Hybridisation of 63 selected amplified loci to the digoxigenin-labelled oligonucleotide probe (TAA)₅ showed that 69.8% of loci from dry pea and 66.6% of loci from lentil hybridised to the probe. Sequencing of amplified products from chickpea with the primer Ta176 demonstrated that one amplicon contained a microsatellite, whereas another amplicon amplified with the same particular STMS primer pair did not. Amplicons produced from lentil and pea with this primer pairs did not contain microsatellite sequences. Results obtained with Tr7, which amplified a PCR product in lentil and chickpea but not in pea, showed that microsatellite sequences were present in chickpea and absent in lentil. Similar results were obtained with Ts35, which produces amplicons in pea and chickpea; but, again, microsatellite sequences were only present in chickpea. We therefore conclude that STMS derived from chickpea could be used to detect variability between other Leguminosae genera, but it is necessary to verify whether homologous loci are revealed.     

Comparison of the heat stress induced variations in DNA methylation between heat-tolerant and heat-sensitive rapeseed seedlings

DNA methylation is responsive to various biotic and abiotic stresses. Heat stress is a serious threat to crop growth and development worldwide. Heat stress results in an array of morphological, physiological and biochemical changes in plants. The relationship between DNA methylation and heat stress in crops is relatively unknown. We investigated the differences in methylation levels and changes in the cytosine methylation patterns in seedlings of two rapeseed genotypes (heat-sensitive and heat-tolerant) under heat stress. Our results revealed that the methylation levels were different between a heat-tolerant genotype and a heat-sensitive one under control conditions. Under heat treatment, methylation increased more in the heat-sensitive genotype than in the heat-tolerant genotype. More DNA demethylation events occurred in the heat-tolerant genotype, while more DNA methylation occurred in the heat-sensitive genotype. A large and diverse set of genes were affected by heat stress via cytosine methylation changes, suggesting that these genes likely play important roles in the response and adaption to heat stress in Brassica napus L. This study indicated that the changes in DNA methylation differed between heat-tolerant and heat-sensitive genotypes of B. napus in response to heat stress, which further illuminates the molecular mechanisms of the adaption to heat stress in B. napus.     

Mechanisms of Resistance to Tobacco Cutworm (Spodoptera litura F.) and their Implications to Screening for Resistance in Groundnut

Summary The Tobacco cut worm (Spodoptera litura Fab.), a polyphagous defoliating insect is a major pest on groundnut in Asia. Screening germplasm for resistance to Spodoptera litura in the field under high infestation revealed significant genotypic variation. Low damage was observed on Mutant (28-2), NC Ac 343, ICGV 86031, R 9227 and TAG 24. In the laboratory rearing of insect, the resistant genotypes, NC Ac 343, Mutant 28-2 and R 9227 affected larval growth and survival, pupal development, adult emergence and fecundity indicating antibiosis as the principal mechanism of resistance. The reduction in larval weight reared on ICGV 86031 could be due to the toughness of leaves. Though the genotype TAG 24 suffered low damage in the field, the larval and pupal development was normal in the laboratory revealing avoidance/non-preference as the mechanism of resistance. Based on the insight gained from the growth and development of the insect on resistant genotypes, the gain in weight (GIW) of the pre-starved larvae was assessed for its suitability in rapid screening. GIW in 24 h by III instar larvae fed with fully expanded II leaf was found suitable in screening for resistance based on antibiosis. The method could be adopted for screening large breeding populations in a short time under laboratory conditions. The resistant genotypes with different mechanisms of resistance could be hybridized to pool the resistant genes for enhancing the level and effectiveness of resistance in the management of the pest.     

Effects of Short‐Term High Temperature on Photosynthesis and Photosystem II Performance in Sorghum

Gas exchange and chlorophyll a fluorescence transient were examined in leaves of sorghum at high temperatures. No changes were found in photosynthetic rate (Pn) and photosystem II (PS II) performance index on absorption base (PI(abs)) at 40 °C for 1 h. But transpiration rate was enhanced significantly, which served as a self‐protection response for dissipating heat. The Pn decreased significantly at 40 °C for 3 h, and the decrease became greater at 45 °C. Decrease in Pn mainly resulted from stomatal limitation at 40 °C for 3 h, whereas it was due to non‐stomatal limitation at 45 °C. Decline in PS II function indicated by the significant decrease in PI(abs), trapped energy flux and electron transport flux were responsible for the decrease in Pn at 45 °C. PS II reaction centre and oxygen‐evolving complex in the donor side were not affected at high temperatures, but electron transport in the acceptor side was sensitive to high temperature. The PS II function recovered completely 1 day after high temperature stress even as high as 45 °C, which is favourable for sorghum to meet the challenge of global warming. However, Pn did not completely recover possibly due to heat‐induced irreversible damage to CO₂ fixation process.