Capturing genetic variability and selection of traits for heat tolerance in a chickpea recombinant inbred line (RIL) population under field conditions

Chickpea is the most important pulse crop globally after dry beans. Climate change and increased cropping intensity are forcing chickpea cultivation to relatively higher temperature environments. To assess the genetic variability and identify heat responsive traits, a set of 296 F_8–9 recombinant inbred lines (RILs) of the cross ICC 4567 (heat sensitive) × ICC 15614 (heat tolerant) was evaluated under field conditions at ICRISAT, Patancheru, India. The experiment was conducted in an alpha lattice design with three replications during the summer seasons of 2013 and 2014 (heat stress environments, average temperature 35 °C and above), and post-rainy season of 2013 (non-stress environment, max. temperature below 30 °C). A two-fold variation for number of filled pods (FPod), total number of seeds (TS), harvest index (HI), percent pod setting (%PodSet) and grain yield (GY) was observed in the RILs under stress environments compared to non-stress environment. A yield penalty ranging from 22.26% (summer 2013) to 33.30% (summer 2014) was recorded in stress environments. Seed mass measured as 100-seed weight (HSW) was the least affected (6 and 7% reduction) trait, while %PodSet was the most affected (45.86 and 44.31% reduction) trait by high temperatures. Mixed model analysis of variance revealed a high genotypic coefficient of variation (GCV) (23.29–30.22%), phenotypic coefficient of variation (PCV) (25.69–32.44%) along with high heritability (80.89–86.89%) for FPod, TS, %PodSet and GY across the heat stress environments. Correlation studies (r = 0.61–0.97) and principal component analysis (PCA) revealed a strong positive association among the traits GY, FPod, VS and %PodSet under stress environments. Path analysis results showed that TS was the major direct and FPod was the major indirect contributors to GY under heat stress environments. Therefore, the traits that are good indicators of high grain yield under heat stress can be used in indirect selection for developing heat tolerant chickpea cultivars. Moreover, the presence of large genetic variation for heat tolerance in the population may provide an opportunity to use the RILs in future-heat tolerance breeding programme in chickpea.     

纤维素降解菌在高温厌氧消化过程中的作用

前期研究表明梭菌属的新株JC3（Clostridiumsp.Strain JC3）是嗜热产甲烷污泥中的主要纤维素降解菌。本项目采用分子生物学技术定量研究了菌株JC3在高温厌氧消化过程中的作用。在含三个小室（室1,室2,室3）的厌氧折流板反应器（9.5 L）中进行纤维素降解实验,试验温度设为55℃。当负荷为2.5 kg COD.m-3d-1,HRT为2天时,80%以上的COD进料都转化为甲烷。采用专门针对菌株JC3的特异性荧光针来研究折流式反应器中污泥样品。结果表明JC3细胞对DAPI染色细胞的比例从低于0.5%（最低检测极限）增加到9.4%（室1）,13.1%（室2）和21.6%（室3）。这表明由荧光原位杂交技术确定的菌株JC3细胞数与滞留污泥的纤维素降解产甲烷活性密切相关。设计了一个针对菌株JC3纤维素酶基因（纤维二糖水解酶A cbh A）的特异探针,并应用到处理固体废弃物如咖啡渣、废纸、垃圾和纤维素等消化污泥中,采用定量PCR技术发现菌株JC3细胞数与高温消化器中污泥的纤维素降解能力密切相关。因此,在高温厌氧消化过程中,菌株JC3对厌氧水解纤维素是非常重要的。     

Suitability of mapped sequence tagged microsatellite site markers for establishing distinctness, uniformity and stability in aromatic rice

At present, testing for distinctness, uniformity and stability (DUS) of crop varieties relies on a set of morphological characters. These characters suffer fromthe limitations of number, interaction with the environment in which the variety grows and subjectivity in decision-making. The potential of DNA-based markers such as sequence tagged microsatellite site (STMS), for establishing DUS merits investigation. In the present study, a set of 55 mapped STMS markers, selected from 12 linkage groups of rice genome, was used to examine distinctness of 23 aromatic rice genotypes including the commercially important Basmati varieties. Forty-one of these markers (74.5%) showed polymorphism between the varieties. The number of alleles per locus ranged from 2–4 with an average of 2.3. The polymorphism information content (PIC) of the markers varied from 0.083 to 0.665 with an average of 0.338. All the varieties could be differentiated from each other at a low probability (0.07×10^-13) of identical match by chance. The marker-based clustering of the varieties corresponded with the known phenotypic classification, thereby providing confidence in the distinctness established by the mapped STMS markers. The utility of these markers to study uniformity and stability was analysed using a commercially important crossbred Basmati rice variety Pusa Basmati 1(IET-10364) that contributes about 40–50% of Basmati rice export from India. Genotyping of twenty individual plants, grown from the nucleus, breeder, foundation, certified and farmer's saved seed samples using all the 55 markers revealed no variation among the plants. These observations suggested that the set of mapped markers employed in this study could be further used for establishing distinctness of aromatic rice varieties and for studying DUS of the important commercial variety Pusa Basmati 1. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetics of nitrate content of lettuce, 1: Analysis of generation means

Summary The genetics of nitrate content in butterhead lettuce (Lactuca sativa L.) was studied using the mean values of five parental genotypes and several generations obtained from crosses between them. One high nitrate parental genotype was chosen and four low nitrate ones. A diallel analysis showed additive genetic effects to be the major source of variation in generation means. Estimates of additive genetic effects differed significantly between experiments, indicating genotype x experiment interactions. Effects of dominance were relatively small. The size and direction of dominance varied between experiments. Reciprocal differences were of very limited size and also varied between experiments. The inheritance of nitrate content in lettuce fitted the additive-dominance genetic model.     

Current status and future strategy in breeding grasspea (Lathyrus sativus)

Efforts in grasspea (Lathyrus sativus) improvement have increased since the development of lines that are very low in the neurotoxin Beta-N-oxalyl-L-alpha-beta-diamino propionic acid (ODAP); also referred to as Beta-N oxalyl-amino-L-alanine (BOAA). Many programs now address several related aspects of improvement simultaneously. These include reduced ODAP concentrations, insect and disease resistance, nitrogen fixation, agronomic practices, fodder and forage production, and components for increased yielding ability. The coordinated, multidisciplinary approach now being applied to the genetic improvement of grasspea should allow the potential of this largely neglected grain legume to be fully realized.     

Efficient In Vitro Shoot-regeneration Systems in Cassava (Manihot esculenta Crantz)

Two different protocols for in vitro regeneration of cassava using zygotic embryos and nodal axillary meristems have been developed. In both cases, buds were regenerated directly from excised explants without an intervening callus phase after a two-step culture procedure. In cotyledonary explants derived from zygotic embryos, prolific shoot formation occurred within 2—3 weeks on MS medium supplemented with 0.5—5 mg/1 BAP alone or in combination with 0.1 mg/1 NAA. Nodal explants with axillary meristems derived from aseptically grown seedlings or stem cuttings were used to initiate a round compact bulb-like structure on MS medium containing 10 mg/1 BAP. These latter structures, when cultured on MS medium supplemented with 0.1 mg/1 NAA, 1 mg/1 BAP and 0.1 mg/1 GA₃, produced multiple shoots. Somatic embryos isolated at the globular/torpedo stage from zygotic embryo explants were also capable of multiple shoot production on medium with 1 mg/1 BAP. Rooting of regenerated shoots exceeded 95 % in phytohormone-free MS medium. No change in their ploidy levels was observed. Therefore, the protocols developed should be of use in the particle gun and Agrobacterium-mediated genetic transformation of cassava.     

The inheritance of chemical phenotype in <Emphasis Type="Italic">Cannabis sativa</Emphasis> L. (III): variation in cannabichromene proportion

The mechanism that controls the proportion of cannabichromene (CBC), a potential pharmaceutical, in the cannabinoid fraction of Cannabis sativa L. is explored. As with tetrahydrocannabinol (THC) and cannabidiol (CBD), CBC is an enzymatic conversion product of the precursor cannabigerol (CBG). CBC is reported to dominate the cannabinoid fraction of juveniles and to decline with maturation. This ontogeny was confirmed in inbred lines with different mature chemotypes. A consistent CBC presence was found in early leaves from a diverse clone collection, suggesting that CBC synthase is encoded by a fixed locus. Morphological variants possessing a ‘prolonged juvenile chemotype’ (PJC), a substantial proportion of CBC persisting up to maturity, are presented. PJC is associated with a reduced presence of floral bracts, bracteoles, and capitate-stalked trichomes. Genetic factors causing these features were independent of the allelic chemotype locus B that was previously postulated and regulates THC and CBD synthesis and CBG accumulation. In contrast to previously described Cannabis chemotypes, the cannabinoid composition of PJCs showed plasticity in that reduced light levels increased the CBC proportion. The ability of PJC plants to enable the production of pharmaceutical raw material with high CBC purity is demonstrated.     

Assessment of partial resistance to powdery mildew in Chinese wheat varieties

Field trials in two cropping seasons and two locations in central China were conducted on 60 Chinese autumn‐sown wheat varieties to assess their partial resistance to powdery mildew. Mean levels of disease severity ranged from close to 0 to more than 90%. The method of inoculation and the location in which trials were conducted affected the relative performance of the varieties, but these effects were much smaller than the main effect of variety. The area under the disease progress curve was highly correlated with final disease severity, but both were poorly correlated with apparent infection rate. Disease severity was regressed against frequencies of virulence in the Blumeria graminis (syn. Erysiphe graminis) f sp. tritici populations in the trial plots. A vertical distance (D) from the mean mildew severity to the fitted line was calculated for each variety and was used to quantify partial resistance. Five of the 60 varieties, ‘Hx8541’, ‘E28547’, ‘Chuan1066’, ‘Zhe88pin6’ and ‘Lin5064’, consistently expressed relatively low levels of disease despite high frequencies of virulence in the pathogen and had consistently high D‐values. They may therefore have good levels of partial resistance.     

Molecular phylogeny in Indian Citrus L. (Rutaceae) inferred through PCR-RFLP and trnL-trnF sequence data of chloroplast DNA

The taxonomy and phylogeny of Indian Citrus is revisited using PCR-RFLP of the trnD-trnT and rbcL-ORF 106 regions as well as sequence data analysis of the trnL-trnF intergenic spacer region of cpDNA. The study was based on 50 accessions of Citrus genotypes, collected from wild, semi-wild and domesticated stocks. Of the 13 restriction enzymes (RE) used for restriction digestion of the polymerase chain reaction (PCR) amplicons, four (Hinf I, Msp I, Alu I, Hae III) generated 47 restriction fragments, of which 24 (51%) were polymorphic. PCR-RFLP data showed a genetic distance ranging from 0 to 0.79 among 50 accessions of Citrus, and a cluster analysis, based on Neighbor-Joining (NJ) method, placed all the accessions in eight major clusters. Analysis of trnL-trnF sequences from 23 representative accessions of Citrus showed a pair-wise sequence divergence rate in the range of 0–0.064. NJ, minimum evolution (ME) and maximum parsimony (MP) analyses of trnL-trnF sequences produced phylogenetic trees, which placed all the 23 accessions in five clusters. PCR-RFLP analysis resulted in a well resolved phylogenetic tree with branches supported by moderate to high bootstrap values, while the trnL-trnF sequence-based trees showed only moderate to low bootstrap support for the internal tree branches, indicating uncertain origin of some Citrus genotypes. This study shows that the trnL-trnF spacer sequence data can detect genetic variation in Indian Citrus genotypes, but the utility of the data in inferring phylogeny at intra and inter-specific levels is limited probably by factors such as hybridization, bud mutations, apomixis and polyploidy. However, PCR-RFLP and trnL-trnF data supported the recognition of C. maxima, C. medica, and C. reticulata as the basal species of edible Citrus.     

Effect of a cytochrome P450 inhibitor on abscisic acid biosynthesis and stomatal regulation in citrus trees in water-stressed conditions

The effect of diniconazole, an inhibitor of cytochrome P450, on drought tolerance was examined in ‘Shiranui’ [(Citrus unshiu Marc. × Citrus unshiu Osbeck) × Citrus reticulate Blanco] trees. Diniconazole treatment increased abscisic acid (ABA) concentration in leaves compared with untreated controls in water-stressed conditions after 20 days of water-stress treatment. Diniconazole significantly decreased the stomatal aperture at 9 h after application compared with untreated controls in water-stressed conditions. The photosynthetic rate decreased in water-stressed conditions; however, regardless of the earlier stomatal closure induced by diniconazole application, the decrease of photosynthetic rate was delayed by the application.