Identification of resistance source in wheat germplasm against spot blotch disease caused by Bipolaris sorokiniana

Four hundred and twenty-two spring wheat germplasm (Triticum aestivum L.) lines belonging to Indian, CIMMYT and Chinese wheat programme were evaluated for their tolerance against natural epiphytotic conditions of spot blotch caused by Bipolaris sorokiniana at the hot spot location, Pusa, Bihar, India. Of the 422 entries screened, none of the genotype showed immunity to the disease, whereas 52 were resistant, 180 moderately susceptible, 171 susceptible and 19 highly susceptible. Indian germplasm lines tended to be more susceptible than lines originated from CIMMYT and China. Chirya 3, Chirya 7 and Mayoor from CIMMYT showed high degree of resistance to the disease both under field and polyhouse conditions. On the basis of the disease severity under field conditions, 20 promising resistant genotypes and 10 highly susceptible lines were isolated for further testing under artificial epiphytotic conditions in polyhouse for genetic analysis and their potential for spot blotch resistance breeding.     

Assessment of genetic diversity among soybean genotypes differing in response to aerial blight (Rhizoctonia solani Kuhn) using SSR markers

Forty‐seven genotypes and one wild relative of soybean, Glycine soja, were screened for resistance against aerial blight under epiphytotic conditions in the field during the Kharif season of two consecutive years viz., 2016 and 2017. Out of the 48 genotypes screened, only 18 genotypes exhibited a moderately resistant response to aerial blight during both the years of study. In order to perform molecular screening of the genotypes for aerial blight resistance, the genomic DNA obtained from the seedlings of the forty‐eight soybean genotypes was subjected to PCR amplification with 12 SSR markers. The SSR markers Satt 119, Sat_076, Satt 433, Satt 281, Satt 277, Satt 245 and Satt 520 were able to clearly amplify different banding pattern for resistant and susceptible genotypes, out of which Satt 433 and Satt 520 were found to exhibit a pattern, highly similar to the results of field screening of the genotypes with respect to resistant and susceptible reaction to the disease. The eighteen soybean genotypes that exhibited moderately resistant reaction to RAB under field conditions during both the years showed a banding pattern similar to resistant check PS‐1583 in the amplification profile produced by the SSR markers. The polymorphism information content (PIC) from the analysis of amplification profile of the SSR markers used in the study, ranged from 0.58 to 0.95. The dendrogram constructed using UPGMA cluster analysis clearly differentiated the resistant and susceptible genotypes of soybean into two separate groups.     

Molecular characterization of urdbean (Vigna mungo) germplasm related to resistance against urdbean leaf crinkle virus

Urdbean (Vigna mungo) is an important pulse crop grown worldwide. Urdbean leaf crinkle virus (ULCV) is a pathogen of urdbean found in Pakistan that causes huge losses in yield. Forty urdbean varieties/lines were screened against the virus under field conditions during spring season 2009. None of the lines appeared to be highly resistant or resistant. On the basis of a 0-5 disease rating scale and disease severity index, genotypes varied significantly in their reaction to ULCV. Four lines (M-6206, IAM-382-15, IAM-133, and Mash-1) were moderately resistant, eight were rated as moderately susceptible, and 21 as susceptible; the remaining seven lines were highly susceptible. RAPD analyses revealed an extensive amount of variation, which could be used for cultivar identification. Genetic differentiation among urdbean genotypes was similar to the field screening data. The varieties 6065-3 and 6206 were highly susceptible and moderately resistant, respectively, to ULCV under field conditions, confirmed by the RAPD analysis. These varieties were the most diverse varieties in the similarity matrix (67.2%), while the varieties IAM-382-9 and 07M003 were the most similar (98.4%). This information will help in the recognition of available resistant germplasms that can resist this disease and will be utilized for urdbean improvement in Pakistan.     

Quantitative trait loci mapping of Cercospora leaf spot resistance in mungbean, <Emphasis Type="Italic">Vigna radiata</Emphasis> (L.) Wilczek

Cercospora leaf spot (CLS) caused by the fungus Cercospora canescens Illis & Martin is a serious disease in mungbean (Vigna radiata (L.) Wilczek), and disease can reduce seed yield by up to 50%. We report here for the first time quantitative trait loci (QTL) mapping for CLS resistance in mungbean. The QTL analysis was conducted using F₂ (KPS1 × V4718) and BC₁F₁ [(KPS1 × V4718) × KPS1] populations developed from crosses between the CLS-resistant mungbean V4718 and CLS-susceptible cultivar Kamphaeng Saen 1 (KPS1). CLS resistance in F₂ populations was evaluated under field conditions during the wet seasons of 2008 and 2009, and resistance in BC₁F₁ was evaluated under field conditions during the wet season in 2008. Seven hundred and fifty-three simple sequence repeat (SSR) markers from various legumes were used to assess polymorphism between KPS1 and V4718. Subsequently, 69 polymorphic markers were analyzed in the F₂ and BC₁F₁ populations. The results of segregation analysis indicated that resistance to CLS is controlled by a single dominant gene, while composite interval mapping consistently identified one major QTL (qCLS) for CLS resistance on linkage group 3 in both F₂ and BC₁F₁ populations. qCLS was located between markers CEDG117 and VR393, and accounted for 65.5–80.53% of the disease score variation depending on seasons and populations. An allele from V4718 increased the resistance. The SSR markers flanking qCLS will facilitate transferral of the CLS resistance allele from V4718 into elite mungbean cultivars.     

Comparison of Resistance to Cotton Leaf Curl Disease (Multan/Burewala) in Gossypium hirsutum L. Varieties and Breeding Lines

Twenty‐four cotton varieties and advance breeding lines were evaluated for their resistance to cotton leaf curl disease (CLCuD; Multan/Burewala) under natural field and in glasshouse conditions for two consecutive years. Resistance was based on symptom expression and disease severity index. All the cotton genotypes exposed to the vector whitefly in the field and artificially infected by grafting manifested a high level of resistance against CLCuD (Multan) with the exception of genotype NIAB‐999 that was moderately resistant. All the test varieties/breeding lines were highly susceptible to CLCuD (Burewala) both in the field and the glasshouse. However, substantial differences were noted between genotypes for disease index under field conditions. Graft inoculation studies showed that all genotypes inoculated with CLCuD (Burewala) developed disease within 9–13 days whereas those graft‐inoculated with CLCuD (Multan) developed symptoms from 15 to 22 days after grafting. Severe reduction occurred in plant morphology, fibre and yield parameters of cotton variety NIAB‐111 following inoculation with CLCuD (Burewala) as compared with CLCuD (Multan).     

Identification of resistance against Fusarium oxysporum f.sp. lini in linseed germplasm

Screening of germplasm/varieties was made to find out the sources of resistance against F. oxysporum f. sp. lini. Screening was conducted on 78 available germplasm/varieties during 2003–2004 and 2004–2005 in rabi season of linseed under natural conditions. Out of total 78 entries, 27 cultures were found to be resistant to disease as the disease incidence in these cultivars were between 0 and 10%. Twenty-three cultivars fell in moderately resistant category with 10.1–25% wilt incidence. Nine genotypes were found moderately susceptible sho'wing 25.1–50% disease incidence, 14 genotypes were found susceptible showing 50.1–75% and 6 genotypes were found highly susceptible to disease (above 75%).     

Molecular mapping for resistance to pea rust caused by <Emphasis Type="Italic">Uromyces fabae</Emphasis> (Pers.) de-Bary

Pea rust caused by Uromyces fabae (Pers.) de-Bary is a major problem in warm humid regions causing huge economic losses. A mapping population of 136 F_6:7 recombinant inbred lines (RILs) derived from the cross between pea genotypes, HUVP 1 (susceptible) and FC 1 (resistant) was evaluated in polyhouse as well as under field conditions during two consecutive years. Infection frequency (IF) and area under disease progress curve (AUDPC) were used for evaluation of rust reaction of the RILs. A linkage map was constructed with 57 polymorphic loci selected from 148 simple sequence repeats (SSRs), 3 sequence tagged sites (STS), and 2 random amplified polymorphic (RAPD) markers covering 634 cM of genetic distance on the seven linkage groups of pea with an average interval length of 11.3 cM. Composite interval mapping (CIM) revealed one major (Qruf) and one minor (Qruf1) QTL for rust resistance on LGVII. The LOD (5.2–15.8) peak for Qruf was flanked by SSR markers, AA505 and AA446 (10.8 cM), explaining 22.2–42.4% and 23.5–58.8% of the total phenotypic variation for IF and AUDPC, respectively. The minor QTL was environment-specific, and it was detected only in the polyhouse (LOD values 4.2 and 4.8). It was flanked by SSR markers, AD146 and AA416 (7.3 cM), and explained 11.2–12.4% of the total phenotypic variation. The major QTL Qruf was consistently identified across all the four environments. Therefore, the SSR markers flanking Qruf would be useful for marker-assisted selection for pea rust (U. fabae) resistance.     

Understanding the inheritance of mungbean yellow mosaic virus (MYMV) resistance in mungbean (<Emphasis Type="Italic">Vigna radiata</Emphasis> L. Wilczek)

Mungbean, Vigna radiata, third in the series of important pulse crops, still suffers from yield loss due to mungbean yellow mosaic disease caused by mungbean yellow mosaic virus (MYMV). Hence, studies on plant-microbe interaction are necessary for understanding the inheritance of resistance. This study concentrated on identification of linked molecular markers for MYMV resistance and to find the genetic inheritance of MYMV resistance in mungbean. A total of 413 germplasm entries in a MYMV hot spot area (Vamban) were subjected to natural field infection and 13 selected resistant lines were subjected to Agrobacterium infection using strains harboring partial genome of two different MYMV isolates, VA221 and VA239. Among the resistant lines, KMG189 showed strain-specific resistance to VA221 and had no symptoms during field trials. Ninety F₂ genotypes were developed from the cross made between KMG189 (MYMV-resistant) and VBN(Gg)2 (MYMV-susceptible), segregated in the Mendelian single cross ratio 3S:1R; susceptibility of all the F₁s to MYMV suggested that the MYMV resistance in mungbean is governed by a single recessive gene. Two SCAR markers CM9 and CM815 were developed through bulk segregant analysis, and the linkage analysis proved CM815 SCAR marker to be linked at 5.56 cM with MYMV resistance gene and SCAR CM9 had nil recombination percentage, suggesting it to be very closely linked to the MYMV resistance gene. SCAR marker CM9 was present in chromosome number 3 of mungbean suggesting novel loci for virus resistance in mungbean. The identified loci can be used for developing varieties resistant to MYMV in mungbean.     

Inheritance of powdery mildew (Erysiphe polygoni DC) resistance in mungbean (Vigna radiata L. Wilczek)

Detached mungbean (Vigna radiata L.Wilczek) leaves were inoculated with a conidial suspension of a local isolate (TI-1) of the powdery mildew pathogen (Erysiphe polygoni DC) under controlled environment conditions. Based on the latent period and severity of the infection, a rating scale of 0–5 was used to classify the host pathogen interactions. Reactions 0, 1 and 2 were considered resistant and referred to as R0, R1 and R2 while 3, 4 and 5 were classified as susceptible (S). RUM lines (resistant to powdery mildew) and their derivatives are crossed with several susceptible (reaction types 3–5) genotypes and the inheritance of the resistance was studied in the F₁, F₂ and F₃ generations. The results showed that powdery mildew resistance in mungbean is governed by two dominant genes designated as Pm-1 and Pm-2. When both Pm-1 and Pm-2 were present, an R0 reaction was observed after inoculation with TI-1. The resistant reaction was R1 when only Pm-1 was present and R2 in the presence of Pm-2. In the absence of both Pm-1 and Pm-2, susceptible reactions 3, 4 and 5 were observed.     

Tolerance as a mechanism of resistance to Thrips palmi in common beans

Tolerance as a mechanism of resistance to the melon thrips, Thrips palmi Karny (Thysanoptera: Thripidae), in common beans, Phaseolus vulgaris L., was evaluated under field and greenhouse conditions. Seven resistant (Brunca, BH‐5, BH‐60, BH‐130, BH‐144, EMP 486, and FEB 115) and five susceptible (PVA 773, EMP 514, BAT 477, APN 18, and RAZ 136) bean genotypes were assessed according to adult and larval populations, visual damage and reproductive adaptation scores, and yield components in field trials. From these genotypes, four resistant (Brunca, BH‐130, EMP 486, and FEB 115) and two susceptible (APN 18 and RAZ 136) genotypes were selected for quantification of proportional plant weight and height increase changes due to thrips infestation in greenhouse tests. Under medium to high thrips infestation in the field, most resistant genotypes tended to have higher reproductive adaptation and lower yield losses, though they did not always suffer less damage, as compared to susceptible genotypes. In the greenhouse, resistant genotypes showed less reduction in plant dry weight and height increase than did some susceptible ones under the same infestation pressure. Results from both field trials and greenhouse tests suggest the possible expression of tolerance as a mechanism of resistance to T. palmi in the resistant genotype EMP 486, and confirm the existence of antixenosis in FEB 115, whereas tolerance might be combined with other resistance mechanisms in Brunca.     

In vitro screening of coffee genotypes for resistance to coffee berry disease (Colletotrichum kahawae)

An in vitro protocol was developed to screen Coffea arabica genotypes for resistance to coffee berry disease caused by Colletotrichum kahawae. Initially, cultural conditions which influenced the growth of isolates of C. kahawae on agar media suitable for callus growth were determined. The growth of the fungus on the callus derived from susceptible and resistant genotypes was then assessed. This ensured that no detrimental competition for nutrients between the pathogen and the calli occurred. Optimisation of the concentration of the phytohormones added to the media, the temperature and incubation period were found to be important in the expression of differential responses of calli to inoculation with the pathogen as detected by measurement of hyphal growth. The screening of calli of nine C. arabica genotypes showed that this method identified genotypes highly resistant or susceptible to the disease and was sufficiently sensitive to distinguish those genotypes with moderate or low resistance.     

Role of antioxidant and anaerobic metabolism enzymes in providing tolerance to maize (Zea mays L.) seedlings against waterlogging

The present investigation was undertaken to identify the possible mode of mechanism that could provide tolerance to maize (Zea mays L.) seedlings under waterlogging. Using cup method, a number of maize genotypes were screened on the basis of survival of the seedlings kept under waterlogging. Two tolerant (LM5 and Parkash) and three susceptible (PMH2, JH3459 and LM14) genotypes were selected for the present study. Activities of antioxidant and ethanolic fermentation enzymes and content of hydrogen peroxide (H2O2), glutathione and ascorbic acid were determined in roots of these genotypes after 72 h of waterlogging. Waterlogging treatment caused decline in activities of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) in all the genotypes. However, only susceptible genotypes showed slight increase in glutathione reductase (GR) activity. Significant reduction in APX/GR ratio in susceptible genotypes might be the cause of their susceptibility to waterlogging. The tolerant seedlings had higher GR activity than susceptible genotypes under unstressed conditions. Stress led to decrease in H202 and increase in glutathione content of both tolerant and susceptible genotypes, but only tolerant genotypes exhibited increase in ascorbic acid under waterlogging conditions. In the tolerant genotypes, all the enzymes of anaerobic metabolism viz. alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH) and pyruvate decarboxylase (PDC) were upregulated under waterlogging, whereas in susceptible genotypes, only ADH was upregulated, suggesting that efficient upregulation of entire anaerobic metabolic machinery is essential for providing tolerance against waterlogging. The study provides a possible mechanism for waterlogging tolerance in maize.     

Influence of cytoplasmic male sterility on expression of different mechanisms of resistance in sorghum to Atherigona soccata (Diptera: Muscidae)

Atherigona soccata (Rondani) (Diptera: Muscidae) is one of the most important pests of sorghum, Sorghum bicolor (L.) Moench, in Asia, Africa, and the Mediterranean Europe. Exploitation of cytoplasmic male sterility (CMS) for hybrid production has resulted in considerable narrowing of the genetic base and may increase the vulnerability of this crop to insect pests. Therefore, we studied the expression of different mechanisms of resistance in sorghum to A. soccata in CMS (A) and maintainer (B) lines of 12 genotypes under field and greenhouse conditions. The CMS lines of A. soccata-resistant genotypes were preferred for oviposition (78.5 versus 71.5% plants with eggs) and suffered greater deadheart incidence (47.6 versus 41.6%) than the corresponding maintainer lines, whereas such differences were not apparent in CMS lines belonging to the susceptible genotypes (92.7 versus 92.3% plants with eggs and 75.6 versus 74.6% deadhearts) under multichoice field conditions. Similar differences also were observed under controlled conditions in the greenhouse. The larval period (9.0 versus 8.8 d) and pupal mortality (18.4 versus 13.4%) were greater on maintainer lines than that on the CMS lines in the resistant group. The male and female pupal weights, fecundity, and antibiosis index were greater on the CMS than on the maintainer lines. The maintainer lines showed better recovery resistance than the CMS lines, but no such differences were observed in tiller deadhearts. The differences in susceptibility to A. soccata were greater in the A. soccata resistant CMS and maintainer lines than in the CMS and maintainer lines belonging to susceptible genotypes. Conversion of A. soccata-resistant genotypes into alternate less susceptible cytoplasmic backgrounds may be undertaken for developing sorghum hybrids with stable resistance to A. soccata.     

Differences in responses to iron deficiency among various cultivars of mungbean (Vigna radiata (L.) Wilczek)

Ten mungbean cultivars were evaluated for their resistance to iron deficiency in view of chlorosis symptoms, plant growth and seed yield under field conditions on a calcareous soil in Thailand. The KPS2 cultivar was highly susceptible; the KPS1, PSU1 and Pag-asa 1 cultivars were somewhat susceptible; the VC1163B cultivar was moderately tolerant; the CN36, CN60, UT1 and CNM-I cultivars were tolerant; and the CNM8509B cultivar was very tolerant to iron deficiency. Foliar application of a solution of 5 g L^-1 ferrous sulphate was effective in correcting chlorosis that was induced by iron deficiency, and it enhanced both the growth and the yield of susceptible cultivars. Compared with the susceptible cultivar KPS2, the tolerant cultivar UT1 had a greater ability to lower the pH of the nutrient solution in response to iron deficiency. The root-associated Fe³⁺-reduction activity of UT1 that had been grown in -Fe medium was similar to that of the plants grown in +Fe medium when the acidification of the medium occurred. Acidification of the medium in response to iron deficiency might contribute to the efficient solubilization of iron from calcareous soils, and it related more closely to the resistance to iron deficiency than Fe³⁺ reduction by roots in mungbean cultivars.     

小麦种质对茎基腐病抗性评价及优异种质筛选

小麦茎基腐病是由镰孢菌侵染引起的一种世界性土传病害,近年来已严重威胁到我国小麦的安全生产。为筛选具有茎基腐病抗性的小麦种质资源,本研究采用孢子悬浮液浸种法,分别以国外抗病材料Sunco和中国品种陕253为抗病和感病对照,对670份我国小麦品种(系)进行了茎基腐病温室苗期抗性鉴定。结果发现,我国供试品种(系)感病材料(病情指数>30)所占比例达到84%,且包含多个近年来小麦生产中的主推品种,表明我国小麦品种总体抗性水平低是导致茎基腐病近年来发病频率与程度不断增加的重要原因之一。经多轮筛选,发掘获得15份抗病表现稳定、抗性水平与抗病对照Sunco相仿的材料。15份材料平均病情指数在10.9~19.4之间,其株高、抽穗期等农艺性状表现出较为丰富的变异,为我国小麦抗茎基腐病品种选育和抗性遗传研究提供了种质资源。     

Screening and Assessment of Selected Chilli (<i>Capsicum annuum</i> L.) Genotypes for Drought Tolerance at Seedling Stage

This study was undertaken to investigate oxidative stress tolerant mechanisms in chilli (Capsicum annuum L.) under drought genotypes through evaluating morphological, physiological, biochemical and stomatal parameters. Twenty genotypes were evaluated for their genetic potential to drought stress tolerant at seedling stage. Thirty days old seedlings were exposed to drought stress induced by stop watering for the following 10 days and rewatering for the following one week as recovery. Based on their survival performance, two tolerant genotypes viz. BD-10906 and BD-109012 and two susceptible genotypes viz. BD-10902 and RT-20 were selected for studying the oxidative stress tolerance mechanism. Drought reduced root and shoot length, dry weight, ratio, petiole weight and leaf area in both tolerant and susceptible genotypes, and a higher reduction was observed in susceptible genotypes. Lower reduction of leaf area and photosynthetic pigments were also found in tolerant genotypes. Moreover, tolerant genotypes showed higher recovery than susceptible genotypes after the removal of stress. A higher reduction of relative water content (RWC) may cause an imbalance between absorbed and transpirated water in susceptible genotypes. Higher accumulation of proline in tolerant genotypes might be helpful to for better osmotic maintenance than that in susceptible genotypes. Tolerant genotypes showed higher antioxidant activity as they showed DPPH radical scavenging percentage than the susceptible genotypes. Moreover, closer stomata in tolerant genotypes than susceptible ones helped to avoid dehydration in tolerant genotypes. Thus, the above morphological, physiological, biochemical and stomatal parameters helped to show better tolerance in chilli under drought stress.     

Screening and identification of random amplified polymorphic DNA (RAPD) markers linked to mungbean yellow mosaic virus (MYMV) resistance in mungbean (Vigna radiata (L.) Wilczek)

Bulk segregant analysis (BSA) and random amplified polymorphic DNA (RAPD) techniques were used to analyse the F2 individuals of susceptible VBN (Gg) 2 × resistant KMG 189 to screen and identify the molecular marker linked to mungbean yellow mosaic virus (MYMV) resistant gene in mungbean. Two DNA bulks namely resistant bulks and susceptible bulks were setup by pooling equal amount of DNA from five randomly selected plants of each disease response. A total of 72 random sequence decamer oligonucleotide primers were used for RAPD analysis. Primer OPBB 05 (5′-GGGCCGAACA-3′) generated OPBB 05 260 fragment in resistant parent and their bulks but not in the susceptible parent and their bulks. Co segregation analysis was performed in resistant and susceptible F2 individuals, it confirmed that OPBB 05 260 marker was tightly linked to mungbean yellow mosaic virus resistant gene in mungbean.     

Effect of heat stress during flowering and pod formation in pea ( Pisum sativum L.)

Heat stress is a major constraint of yield in grain legumes including peas. Increasing global warming and human population now urge to develop climate resilient varieties. The present experiment was conducted over 2 years to evaluate the heat tolerance of 211 pea genotypes. In the present study, the field pea genotypes showed a wide variation for reproductive stage heat stress (RSHS) quantitative traits. Significant positive correlations were found between no. of seeds per plant and no. of pods per plant; seed diameter (mm) and 25-seed weight (g) in heat tolerant as well as heat susceptible genotypes. Principal component analysis revealed two major principal components contributed approximately 91% of total variations and heat tolerant and susceptible genotypes separately formed two major clusters. Stepwise multiple regression analysis revealed that no. of seeds per plant was the best predictor for no. of pods per plant. On the basis of four RSHS traits, the most prominent heat tolerant pea genotypes identified in the present study JP-625, IARI-2877, PMR-38 II, EC-318760, EC-328758 and IARI-2904 would better combat RSHS and provide yield stability under changing climatic conditions.     

Effect of Saline Environment on Yield and Smut Disease Severity of Different Corn Genotypes (Zea mays L.)

The effects of saline irrigation water on growth, yield, smut index and leaf CI content were investigated under field conditions. The plants were grown in 1994 and 1995 and artificially infected with U. maydis- spore-suspension at V₆ and V₁₀ leaf stages. The severity was rated on a 1–9 scale, whenever the symptotns appeared. The results showed that superior growth, greater yield performance, associated with lower smut incidence and leaf Cl contents were generally observed on Taba than its counterparts. Increasing salinity exhibited significant increases in leaf CI and induced marked inhibition on growth, yield and SI data. The salt sensitivity of corn genotypes, as revealed from the comparison of the slope of linear regression equations, was related to 1.00: 1.19;1.42 for cvs Taba, 310 and 320, respectively. The differential yield response with respect to SI data proved that smut susceptibility was inversely related to salt tolerance concept. The data of Cl-disease interaction showed that smut reactions of highly susceptible and resistant genotypes (cvs 320 and Taba) were hardly monitored by plant Cl content, revealing that disease resistance is genetically controlled. Conversely, the progressive modifications of smut reaction, accompanied by CI accumulation in the moderately susceptible genotype 310 proved that disease resistance was environmentally conditioned by salt stress.