AFLP Fingerprinting in Pigeonpea (Cajanus cajan (L.) Millsp.) and its Wild Relatives

Detection of DNA polymorphism in cultivated pigeonpea (Cajanus cajan) and two of its wild relatives Cajanus volubilis and Rhynchosia bracteata is reported here for the first time using amplified fragment length polymorphism (AFLP) fingerprinting. For this purpose, two EcoRI (three selective nucleotides) and 14 MseI (three selective nucleotides) primers were used. The two wild species shared only 7.15% bands with the pigeonpea cultivars, whereas 86.71% common bands were seen among cultivars. Similarly, 62.08% bands were polymorphic between C. volubilis and pigeonpea cultivars in comparison to 63.33% polymorphic bands between R. bracteata and pigeonpea cultivars, and 13.28% polymorphic bands among pigeonpea cultivars. The cluster analysis revealed low polymorphism among pigeonpea cultivars and very high polymorphism between cultivated pigeonpea and its wild relatives. The AFLP analysis also indicated that only one primer combination (EcoRI + ACT and MseI + CTG), at the most any four primer pair combinations, are sufficient for obtaining reliable estimation of genetic diversity in closely related cultivars like pigeonpea material analyzed herein. AFLP analysis may prove to be a useful tool for molecular characterization of pigeonpea cultivars and its wild relatives and for possible use in genome mapping.     

Impact of nodule damage by Rivellia angulata on N2-fixation,growth, and yield of pigeonpea (Cajanus cajan L. Millsp.) grown in a Vertisol

Summary Damage caused by Rivellia angulata larvae to pigeonpea root nodules at the ICRISAT center in India was greater in the crop grown on Vertisols (up to 86%) compared to that on Alfisols (20%). Attempts to quantify the field effects of nodule damage on growth and yield of pigeonpea in a Vertisol, involving many heavy applications of soil insecticides (aldrin and hexachlorocyclohexane) failed because the insecticides did not control the pest and adversely affected the growth of the pigeonpea and the subsequent crop of sorghum (Sorgorum bicolor L. Moench). The impact of nodule damage on pigeonpea growth, yield and nutrient uptake was successfully studied in greenhouse-grown plants at three N levels. In this pot study, artificial inoculation with Rivellia sp. led to substantial nodule damage (70%). The results of this damage were a significant overall reduction in nodule dry weight (46%), acetylene reduction activity (31%), total leaf area (36%), chlorophyll content of leaves (39%) and shoot dry weight (23%) 68 days after sowing. At maturity, Rivellia sp. infestation caused significant reductions in top dry weight (22%), root and nodule dry weight (27%), seed dry weight (14%), and total N (29%) and P uptake (19%). The problems and prospects of manipulating nodule damage so as to reduce N losses in pigeonpea are discussed.Submitted as JA No. 756 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT)     

Variation of morpho-physiological traits in geographically diverse pigeonpea [Cajanus cajan (L.) Millsp] germplasm under different phosphorus conditions

Genotypic variation for morpho-physiological parameters, phosphorus (P) content and root acid phosphatase activity was studied in 52 pigeonpea genotypes. Data related to shoot (length, dry weight, number of leaves, and leaf area), root (volume, length, dry weight, area, perimeter, and number of root tips), acid phosphatase activity, and P content (root, stem, and leaf) were recorded at 60 days after sowing (DAS). The P use efficient genotypes showed high root length, root area, root perimeter, root dry weight, P content in leaves, and root to shoot dry weight ratio under the P not added condition. Significant variation was found among genotypes for root- and shoot-associated characteristics under both P treatments. The P use efficient genotypes with improved root morphological phenes have potential to acquire and utilize more P from immobile soil bound P sources may be of additional factor for increasing efficiency of acquisition and utilization of supplied P fertilizer.     

Effect of temperature on flavonoid production in pigeonpea [Cajanus cajan (L) Millsp] in relation to nodulation

To ascertain the reasons for poor nodulation of pigeonpea, we studied the effect of high temperature on the production of flavonoids by the pigeonpea host. A high temperature affected flavonoid production by pigeonpea and mungbean. At 37°C pigeonpea root exudates contained four flavonoids and the root extract contained five. The proportion of the second flavonoid in the pigeonpea and the mungbean was higher and the proportion of the third flavonoid was lower at 37°C compared to 30°C. At the higher temperature the flavonoids exuded from pigeonpea roots were same those in the root homogenate.     

Effect of high temperature on plasmid curing of Rhizobium spp. in relation to nodulation of pigeonpea [Cajanus cajan (L.) Millsp.]

Summary Fifty-six isolates of Rhizobium and Bradyrhizobium spp. (Cajanus) were studied for their plasmid profile and N₂-fixation efficacy. One to three plasmids were reproducibly detected in all the Rhizobium spp. strains but no plasmid was detected in the Bradyrhizobium spp. strains. Rhizobium sp. strain P-1 was mutagenized by Tn5 and three nod^– and six nod⁺fix^– were screened for symbiotic parameters. Neomycin-sensitive mutants were isolated by elevated temperatrue (40°C) from tranconjugants carrying Tn5 insertions. The high temperature cured these mutants from the single large plasmid present in the parent strain P-1. All these cured mutants were nod^–, indicating that the genes for nodulation were present on this plasmid, which is readily cured at a high temperature (40°C). The high temperature in the semi-arid zones of Haryana could be responsible for the low nodulation of pigeonpea because the plasmid carrying the nodulation genes is cured at 40°–45°C giving rise to non-nodulating mutants.     

转GhB301基因棉花响应枯萎病菌侵染的转录组分析

乙烯响应因子(ethylene responsive factor, ERF)可以激活或者抑制下游病程相关蛋白基因的表达,在植物抗病信号转导途径中发挥着重要作用。为探究ERF-B3亚组基因GhB301在棉花抗枯萎病中的分子调控机制,本研究利用已获得的转GhB301基因棉花株系,通过孢子悬浮液蘸根的方法对转GhB301基因棉花株系(N)和野生型对照(WT)进行接菌处理,抗病性鉴定结果表明,过表达GhB301的N株系增强了对枯萎病的抗病性,其病情指数为14.77,显著低于WT(病情指数为37.50);枯萎病菌侵染0、6、12、24、48 h后,利用RNA-seq技术对N和WT的根部组织进行测序分析,共得到273 111 170个clean reads,其Q30均大于87.64%,将clean reads与陆地棉参考基因组TM-1比对,获得了14 021个差异表达基因(DEGs)。与WT相比,转基因株系能够在病原菌侵染后更快速地做出响应。GO及KEGG富集分析发现共有135个DEGs参与氧化还原过程,67个DEGs参与防御反应,31个DEGs参与苯丙烷类生物合成,推测这些DEGs可能与转基因棉花的枯萎病抗性增强存在密切关系。本研究结果为阐明GhB301基因响应棉花枯萎病菌侵染规律奠定了基础。     

Patterns of diversity in pigeonpea (<Emphasis Type="Italic">Cajanus cajan</Emphasis> (L.) Millsp.) germplasm collected from different elevations in Kenya

Pigeonpea germplasm accessions collected from low (<500 m), medium (501–1000 m), high (1001–1500 m) and very high elevation zones (>1500 m) of Kenya were evaluated for 15 agronomic traits and seed protein content at ICRISAT, Patancheru, India. There were significant differences (P < 0.001) among elevation zones for the number of primary and secondary branches, days to 75% maturity, pod length, seeds per pod, 100-seed weight and seed yield. Mean values indicated that the accessions from low elevation zone were significantly different from those collected in higher elevation zones for early flowering and maturity, number of primary branches, pod length, number of pods per plant, seeds per pod, 100-seed weight, seed yield and harvest index. None of the accessions collected in Kenya belonged to extra early (<80 days to 50% flowering) and early (80–100 days to 50% flowering) maturity groups, as defined by time to flowering at Patancheru, India. Mean diversity index based on all characters indicated that accessions from the low elevation zone are more diverse than those from the higher elevation zones. Frequency distribution for trait extremes indicated that the accessions from the low elevation zone were early to flower and mature, short statured, produced more primary and secondary branches with high pod bearing length, long pods, more pods per plant, more seeds per pod, a high seed yield and harvest index. Accessions from the very high elevation zone were late flowering, with a large number of tertiary branches, large seeds and a high shelling percentage and could be a source for cold tolerance and the breeding of vegetable types. Results suggest that the elevation of collection sites is therefore a very important determinant of variation patterns of pigeonpea in Kenya.     

Karyotypic and genome size variation in Cajanus cajan (L.) Millsp. (pigeonpea) and some wild relatives

Detailed karyotypes and 4C DNA amounts have been studied in five cultivars of Cajanus cajan and 20 species belonging to Cajanus, Rhynchosia, Dunbaria, Flemingia and Paracalyx. C. cajan shows intraspecific variability and its karyotype is most similar to that of C. cajanifolius (sect. Cajanus) in the morphology and number of satellite chromosomes and the lack of any correlation between chromosome size within the complement and asymmetry. Karyotypes of C. lineatus and C. sericeus belonging to sect. Atylia are similar with respect to maximum r-index and the ratio of longest and shortest chromosomes in their respective complements. C. acutifolius (sect. Frutocosa) is distinct in having a very low ratio (1.44) between the longest and the shortest chromosomes, while in C. albicans, C. goensis, C. scarabaeoides (sect. Cantharospermum) there is no chromosome pair with r-index >2.0. C. mollis and C. volubilis (sect. Volubilis) show similarity with regard to ratio between the longest and the shortest chromosomes and a distinct chromosome pair with an arm ratio of 2.7 and C. platycarpus is distinct (sect. Rhynchosoides) in having the smallest ratio (1.36) between the longest and the shortest chromosomes. 4C DNA amounts in Cajanus and Rhynchosia vary between 3.28 pg to 11.69 pg and 4.92 pg to 11.13 pg respectively, while in Dunbaria, Flemingia and Paracalyx these vary between 4.70 and 7:06 pg. In the genus Cajanus karyotypic features and 4C DNA amounts agree with the sectional classification and bring out a close relationship between C. cajan and C. cajanifolius. This is supported by studies based on seed protein patterns, isozyme analysis, trypsin and chymotrypsin inhibitor patterns, RFLP and RAPD data and crossability relationships. A clear relation between DNA amount and abnormalities effecting the extent of pairing and recombination in interspecific hybrids shows the importance of this study in developing future breeding programme involving C. cajan and its wild relatives that are of potential value.     

Phylogenetic relationships of Cajanus cajan (L.) Millsp. (pigeonpea) and its wild relatives based on seed protein profiles

Total seed storage protein of 9 accessions of cultivated C. cajan and 10 wild Cajanus species was reported and compared by sodium dodecyl sulphate polyacrylamide gel electrophoresis. A considerable variation was detected among the protein profiles of different accessions of C. cajan while those of wild species were very specific and distinctly different from each other. Relative similarities between various taxa were estimated by Jaccard's similarity index and cluster analysis was performed to produce a UPGMA dendrogram. The clustering of 10 wild species and C. cajan more or less agrees with their sectional classification and available data based on morphological characteristics, crossability, genome pairing in hybrids and nuclear RFLPs. The species closest to C. cajan is C. cajanifolia although the accessions of C. cajan also share some bands present in the profiles of C. scarabaeoides, C. goensis, C. lineatus, C. acutifolius and C. volubilis. This points towards polyphyletic origin of the cultigen which has been discussed in this paper.     

有机肥与生防菌结合防治香蕉枯萎病的初步研究

研究在适宜化肥用量条件下配施有机肥与生防菌、土壤消毒与生防菌对香蕉植株生长和香蕉枯萎病防治效果的影响。结果表明,两个配施鸡粪处理分别在香蕉营养生长期和抽蕾期蕉株最高,茎围最大。鸡粪淋施复合芽孢杆菌处理成熟蕉果可溶性糖含量与Vc含量最高,产量最高,比氮磷钾处理增产8.9%;在种植后5个月才开始发病,发病时间延缓,发病后病况稳定且发病率最低,为17.8%,比其它处理降低6.6～46.6个百分点,防治效果最好。受香蕉收获株数减少、种植成本上涨、香蕉价格不稳定等因素影响,建议在已发枯萎病蕉园种植其它作物以获得更高种植效益。     

Seedling characteristics and retention of current photosynthates in leaves in relation to initial growth in pigeonpea (Cajanus cajan L. Millsp.) and cowpea (Vigna sinensis Endl.)

Improved varieties of pigeonpea (Cajanus cajan L. Millsp.) still retain some growth characteristics of perennial types of wild species such as slow seedling growth and rato on ability which are considered to be major yield-restricting characteristics when the growth period is shortened. Studies on the physiological background of slow initial growth may help improve the potential productivity of short-duration pigeonpea. It was found that the low seedling vigor of pigeonpea could be ascribed to the small seed size (Narayanan et al. 1981; Brakke and Gardner 1987), low rate of photosynthetic CO₂ fixation (Rawson and Constable 1981), and low rates of metabolic processes (Huber and Hanson 1992; Ito et al. 1996). However, few authors have related the slow growth to the retention of current photosynthates and leaf respiration during the night. In contrast, seedling growth of cowpea (Vigna sinensis Endl.) was reported to be vigorous (Brakke and Gardner 1987).

The objectives of this study were to characterize the initial growth of pigeonpea in comparison with cowpea. Two experiments were carried out, one dealing with growth analysis and the other with O2 uptake of leaf and retention of current photosynthates during the night.     

苦瓜产量杂种优势与配合力研究

选用8个高世代绿苦瓜自交系,按Griffing完全双列杂交第2种方法设计试验,对苦瓜产量的杂种优势和配合力效应进行研究。结果表明:28个杂交组合有26个呈现正向离中优势,有23个呈现正向超亲优势,22个组合比对照增产;在8个试验亲本中,4个亲本的一般配合力效应为正向效应,另外4个为负向效应,亲本B的一般配合效应值极显著高于其它亲本;21个组合双亲间特殊配合力效应表现为正向效应,效应值最高的组合为E×H(2.15),其次是B×H(1.51)、D×H(1.28)、A×G(1.23)和C×G(1.05)。并对苦瓜丰产性育种的亲本选择选配进行了讨论。     

Combining ability of yield and yield components among Fusarium oxysporum f.sp. strigae-compatible and Striga-resistant sorghum genotypes

ABSTRACT

Use of sorghum [Sorghum bicolor (L.) Moench] cultivars with partial resistance to Striga spp. and Fusarium oxysporum f.sp. strigae (FOS) represents a novel strategy to control Striga. This study aimed to identify the nature of gene action controlling grain yield and yield components and to select promising sorghum crosses possessing both FOS compatibility and Striga resistance, along with good combining ability effects. One-hundred hybrids, developed from pairwise matings among 10 FOS compatible, high-yielding female lines and 10 Striga-resistant male lines, were evaluated with and without FOS inoculation. The F₁s were field evaluated at three locations in Tanzania known for their severe Striga infestation, using an alpha lattice design with two replications. General (GCA) and specific combining ability (SCA) variances were significant for grain yield per plant, hundred-seed weight, plant height, flowering time and the number of Striga plants. The study demonstrated FOS inoculation to be an effective means of controlling Striga. Families 675?×?672, AS435?×?3993 and 4643?×?AS436 displaying large SCA effects for grain yield, and 4567?×?AS429, 3424?×?AS430 and 3424?×?AS436 with small SCA effects for Striga counts should be useful genetic resources for breeding and integrated Striga management.     

沼液处理对连作西瓜枯萎病发生、产量及品质的影响

利用沼液防治土传病害是实现沼液高值利用重要途径。以西瓜优佳018为材料,采用田间试验方式,研究等氮磷钾养分下,沼液结合覆膜、闷棚等综合防治措施,对西瓜连作土壤中尖孢镰刀菌数量的削减效果以及对西瓜产量及品质的影响。结果表明:水或沼液结合覆膜、闷棚等处理连作土壤20天,能有效降低土壤中尖孢镰刀菌的数量,其中沼液处理降低幅度更大。处理期间,沼液和水处理土壤温度、湿度显著高于对照。沼液处理减缓了西瓜枯萎病大面积发生的时间,前期(西瓜移栽后的45天内)西瓜枯萎病发病率和病情指数均显著低于水处理和对照,开花期之后发病率大幅提高,因而西瓜收获时沼液处理与对照发病率无显著差异,但病情指数显著低于对照。与对照和水处理相比,沼液处理西瓜产量、商品果产量、维生素C含量均显著提高,而硝酸盐含量降低,但沼液处理西瓜含糖量低于对照。以上表明,利用沼液处理西瓜连作土壤能有效增加西瓜产量,提升品质,并对枯萎病的发生有一定的延缓作用。     

番茄枯萎病对植株维管束危害及抗氧化系统影响的研究

为了在早期诊断和确定番茄枯萎病的发生, 本文采用溶液培养方法研究了番茄(Lycopersicon esculentum Miller)幼苗剪根接种不同浓度枯萎病菌后染病植株维管束受害程度和抗氧化系统的响应。试验设4个病原菌梯度处理, B1(10⁴ cfu·mL^-1)、B2(10⁶ cfu·mL^-1)、B3(10⁷ cfu·mL^-1)、B4(10⁸ cfu·mL^-1), 以不接病原菌为对照; 分别在接种病原菌后4 d、8 d、12 d、16 d、20 d测定维管束褐变情况和抗氧化系统的变化。结果表明, 在水培条件下, 接种病原菌16 d植株维管束出现褐变, 其受害程度随病原菌接种浓度提高而增大; 维管束中病原菌只在B4处理中有检出。番茄叶片中丙二醛(MDA)含量随接种时间呈先降后升趋势, 12 d开始逐渐上升, 20 d达到最高, 各接菌处理均显著高于对照, 且B4处理显著高于其他处理; 过氧化物酶(POD)活性先缓慢下降, 12 d后回升, B4则急剧上升; 多酚氧化酶(PPO)活性逐渐上升, 接菌16 d时达到高峰; 随接菌浓度的提高, MDA含量、POD和PPO活性均有所增加, 尤以接菌浓度为10⁸ cfu·mL^-1时3种指标显著高于其他处理, 分别是未接菌植株的13.1倍、12.9倍和1.9倍; 而培养时间对番茄叶片中过氧化氢酶(CAT)活性的影响没有明显规律, 对照菌株CAT活性显著高于各接菌处理, 说明CAT活性对番茄枯萎病病原菌没有响应。本研究结果表明, 结合番茄茎的维管束褐变现象, 认为番茄叶片中MDA含量、POD和PPO活性可作为早期判断番茄是否感染枯萎病的重要指标。     

AFLP analysis in pigeonpea (<Emphasis Type="Italic">Cajanus cajan</Emphasis> (L.) Millsp.) revealed close relationship of cultivated genotypes with some of its wild relatives

A total of 561 amplified fragment length polymorphism (AFLP) loci were generated and used to study the genetic diversity of wild and cultivated genotypes of pigeonpea. Out of 561 marker loci, 558 were polymorphic with an average of 76.12 bands. Analysis of molecular variation (AMOVA) revealed significant strong population structure when genotypes were structured according to continent of origin (F_ST = 0.22) also when structured into cultivated and wild genotypes (F_ST = 0.16). Maximum polymorphic loci were observed in cultivated species C. cajan (352) which is due to more number of genotypes used while, a minimum number of 45 polymorphic bands were obtained in C. acutifolius. Highest (0.291) average gene diversity was recorded in species C. mollis and lowest (0.079) average gene diversity was recorded in C. acutifolius. The 33 genotypes grouped into 14 clusters at 26% Jaccard’s similarity coefficients. Clustering analysis revealed most cultivated genotypes grouped into one major cluster while, the wild genotypes grouped into many clusters at 26% Jaccard’s similarity revealing greater diversity within wild species as compared to cultivated genotypes. From among the various cultivated genotypes studied, four genotypes BRG 3, ICP 7035, TTB 7 and ICP 8863 were molecularly and morphologically diverse and were used as parental genotypes to study nature of inheritance and to identify markers linked to sterility mosaic disease.     

Differences between Cajanus cajan (L.) Millspaugh and C. cajanifolius (Haines) van der Maesen, the progenitor species of pigeonpea

A study was undertaken to know the difference/diversity between pigeonpea and its closely related wild species C. cajanifolius by studying their morphology, crossability, cytology of the hybrid between the two, and molecular studies. Studies revealed that there are at least 5–6 traits that separate the two species such as flower morphology, pod color and morphology, pod constriction, seed color and strophiole, 100 seed weight that separate C. cajan from C. cajanifolius. Molecular studies revealed that a genetic dissimilarity index value ranging from 0.81 to 0.94 exists between the two species.     

Use of GGEbiplot methodology and Griffing's diallel method for genetic analysis of partial resistance to phoma black stem disease in sunflower

Abstract

The objectives of the present study were to estimate the general combining ability (GCA) and specific combining ability (SCA) for partial resistance to phoma black stem and to identify the most promising combination for the selection of improved breeding lines. The response of five parental genotypes and their F1 hybrids to a phoma black stem isolate (MA6) were evaluated in a diallel programme under controlled growth chamber conditions. Significant GCA and SCA indicate that both additive and non-additive gene effects contributed in the inheritance of partial resistance to phoma black stem, however, the Baker ratio showed that the additive genetic effects were more important than nonadditive ones. It is recommended that the GGEbiplot methodology could be an excellent tool for visualizing entry by tester (diallel) data. By using this technique to analyse black stem severity data, interaction among the sunflower genotypes in providing partial resistance to phoma black stem was clearly identified. Based on GGEbiplot presentation and Griffing's diallel analysis, the mutant line ‘M6-54-1’ showed the largest GCA, indicating contribution towards partial resistance, and the genotype B454/03 presented the smallest GCA, indicating contribution towards susceptibility. Our results show that the F1 hybrids ‘SDR18×B454/03’ and ‘M6-54-1×B454/03’ showing heterosis for partial resistance to phoma black stem come from the crosses between a susceptible genotype ‘B454/03’ and two partially resistant genotypes (SDR18 and M6-54-1), originated from different breeding programmes. We conclude therefore that these genotypes possess at least some different resistance genes, which were expressed in the hybrids and led to the observed effects.