Variation in the Agronomic and Morphological Traits of Iranian Chickpea Accessions

Landraces of chickpea (Cicer arietinum L.) in Iran have not been adequately characterized for their agronomic and morphological traits. Such characterization would be helpful in the development of improved cultivars, so in this study 362 chickpea accessions, collected from the major chickpea growing areas of Iran, were evaluated to determine their phenotypic diversity. High coefficients of variation (CVs)were recorded in pods/branch, seeds/pod, yield/plant, seeds/plant, pods/plant and branches/plant. Using principal component (PC) analysis, the first four PCs with eigenvalues more than 1 contributed 84.10% of the variability among accessions, whereas PC5 to PC10 were less than unity. PC1 was positively related to days to first maturity, days to 50% flowering and days to 50% maturity. The characters with the greatest weight on PC2 were seeds/plant and yield/plant, whereas PC3 was mainly related to pods/plant, seeds/pod and 100-seed weight, and PC4 was positively related to pods/branch and negatively to branches/plant. The germplasm was grouped into four clusters using cluster analysis. Each cluster had some specific characteristics of its own and the cluster I was clearly separated from clusters Ⅱ, Ⅲ and Ⅳ. These accessions are an important resource for the establishment of a core collection of chickpeas in the world.     

鹰嘴豆种质资源农艺性状遗传多样性分析

以100份鹰嘴豆种质资源为材料,应用聚类分析和主成分分析方法,对15个主要农艺性状的遗传多样性进行分析。结果表明,参试材料存在广泛的遗传多样性。其中,多样性指数最高的是株高,其次是百粒重;性状变异系数最大的是单株荚数,其次是单株粒重;基于各种质间形态标记的遗传差异,将100份鹰嘴豆种质聚类并划分为4大类群。第Ⅰ类群可作为选育丰产中粒型和株高适中的品种,第Ⅱ类群可作为选育矮秆耐密及特异粒色(型)品种,第Ⅲ类群丰产性较差可作为选育子粒球型、光滑的品种,第Ⅳ类群可作为选育大粒型、适宜机械化收获的品种。9个数量性状的主成分分析结果表明,前4个主成分累计贡献率达73.91%,各主成分性状载荷值反映了主要数量性状的育种选择潜力。综合分析种质资源农艺性状,为鹰嘴豆的有效利用提供一定的科学依据。     

Restriction Fragment Length Polymorphism and Random Amplified Polymorphic DNA Analysis of Chickpea Accessions

Genetic diversity analysis was carried out in chickpea accessions using restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) techniques. RFLP analysis using 26 Pst I sub-genomic clones on ten chickpea accessions in 130 probe-enzyme combinations detected polymorphism with only two clones. Pst I clones, CG 141 detected polymorphism in ICC 4918 and Pusa 209 while CG 500 detected polymorphism in Pusa 261, ILC 26 and in ILC 13326. These clones detected very few polymorphic markers. Analysis using 10 Eco RI clones on twelve chickpea accessions have shown better hybridisation signal and one clone detected polymorphism in Pusa 256. RFLP analysis of both cultivated and wild Cicer species using heterologous DNA probe Cab3C revealed polymorphism only in wild Cicer species (Cicer reticulatum L., JM 2100). RAPD analysis of 13 chickpea accessions which includes mutants of C 235 and E100Y showed greater degree of polymorphism with 1 - 5 unique DNA bands for all the accessions. Phylogenetic analysis of the RAPD data helped to group the accessions. C 235 and its mutants were found to be closely grouped while E100Y and its mutant E100Ym grouped apart. Desi and kabuli chickpea accessions however, could not be separately grouped. This revised version was published online in July 2006 with corrections to the Cover Date.     

小豆种质资源农艺性状综合鉴定与评价

通过田间观测和室内分析,对来自全国各生态区的262份小豆优异种质资源连续2年进行了农艺性状鉴定及综合评价。结果表明:262份小豆微核心种质资源在考察的16个形态学性状中均具有丰富的遗传变异类型;不同年份间生态环境尤其是光照条件对小豆农艺性状影响较大;主成分分析确定了3类影响因子,表明小豆资源的选育要集中在生长势良好(生育日数较短、株高较矮),单株荚数和单荚粒数多的地方品种;聚类分析结果显示在相似系数为0.40时可将参试材料分为5大类群,各类群间性状差异明显;群体性状表现与地理来源之间没有直接的关系。     

鹰嘴豆种子胰蛋白酶抑制剂的分离纯化与鉴定

为了寻找具有药物作用的天然胰蛋白酶抑制物,采用硫酸铵分级沉淀、离子交换层析(DEAE-纤维素52)及Sephadex G-100凝胶层析等方法, 从鹰嘴豆种子中分离出一种鹰嘴豆胰蛋白酶抑制剂（CPTI）. 研究表明：CPTI对胰蛋白酶有较强的抑制作用,抑制率达80%,而对胰凝乳蛋白酶抑制作用较弱,抑制率为32%, 对胃蛋白酶、木瓜蛋白酶及枯草杆菌蛋白酶均无抑制作用; 用SDS-PAGE测得CPTI近似分子质量为25.7 kD; CPTI具有较高的热稳定性,在100 ℃下加热60 min,对胰蛋白酶活性仍保持78%抑制率; Lineveaer-Burk作图得知该抑制剂属竞争性抑制类型. 动力学测定显示,来自鹰嘴豆中的CPTI对胰蛋白酶的抑制作用常数(Ki)为3.99×10^-7 mol/L.     

Chickpea genotypes differ in their sensitivity to Zn deficiency

Zinc (Zn) deficiency is common in most of the chickpea growing areas of the world and growing Zn-efficient genotypes on Zn-deficient soil is a benign approach of universal interest. Response of 13 chickpea genotypes (10 desi and 3 kabuli) to Zn nutrition was studied in a pot experiment under glasshouse conditions. Plants were grown in a Zn-deficient siliceous sand for 6 weeks and fertilized with 0 (Zn–) and 2.5 mg Zn per kg soil (Zn+). When grown with no added Zn, Zn deficiency symptoms (chlorosis of younger leaves and stipules followed by necrosis of leaf margins) appeared 3–4 weeks after planting and were more apparent in cultivars Tyson, Amethyst and Dooen than Kaniva and T-1587. Zn deficiency reduced shoot growth, but it was less affected in breeding lines T-1587 and CTS 11308 than cultivars Tyson, Dooen, Amethyst and Barwon. Among all genotypes, Tyson produced the lowest root dry weight in Zn– treatment. Zinc efficiency based on shoot dry weight showed marked differences among genotypes; breeding lines CTS-60543, CTS-11308 and T-1587 were 2-fold more Zn-efficient than cultivars Tyson and Dooen. A higher Zn accumulation per plant and higher Zn uptake per g. of root dry weight were recorded in T-1587 and CTS-11308 when compared with Tyson. Root:shoot ratio was increased and proportionally more Zn was transported to the shoot when the soil was deficient. Cultivars that were very sensitive to Zn deficiency tended to have their root:shoot ratio increased by Zn deficiency more than less sensitive cultivars. The insensitive lines T-1587 and CTS-11308 transported more than 70% of the total absorbed Zn to the shoot. It is concluded that chickpea genotypes vary in their sensitivity to Zn deficiency. Advanced breeding lines T-1587 and CTS-11308 are relatively more Zn-efficient compared with Australian chickpea cultivar Tyson. Zn efficiency in chickpea genotypes is probably related to an efficient Zn absorption coupled with a better root to shoot transport.     

Influence of inoculation with Ascochyta rabiei on the mineral contents of resistant and susceptible cultivars of chickpea

The determination of mineral contents of healthy as well as Ascochyta rabiei inoculated resistant, moderately resistant, moderately susceptible and susceptible cultivars revealed that the amount of N, P, Zn and Fe did not vary much in healthy plants of the resistant and susceptible cultivars. The amount of K and S was greater in the susceptible cultivars compared to the resistant cultivars while the reverse was true for Cu and Mn. Barring the recovery of Cu and Fe, the amount of all other elements (N, P, K, S, Zn and Mn) was enhanced upon inoculation of resistant, moderately resistant, moderately susceptible and susceptible cultivars. There was a noticeable increase in the amount of K in the resistant cultivars and the reverse was true for P, S and Mg contents after inoculation.     

Evolution of Ethylene and Methane in Relation to Somatic Embryogenesis in Chickpea

In four genotypes of chickpea (Cicer arietinum L.) BG 362, BG 372, BG 329 and C235 the relationship between somatic embryogenesis of leaf explants and ethylene and methane evolution was studied. In BG 362, which was more embryogenic than other genotypes, a higher ethylene:methane ratio of 5.8:1 at day one after inoculation in the induction medium and a lower ethylene:methane ratio of 2.89:1 in the maturation medium was found. On the contrary, in BG 372 with the least embryogenic potential, a lower ethylene:methane ratio of 1.7:1 in the induction medium and a higher ethylene:methane ratio of 4:1 in the maturation medium was found. Thus, these ratios in induction and maturation stages seems to be markers for embryogenesis in leaf explants of chickpea. This revised version was published online in July 2006 with corrections to the Cover Date.     

基于表型性状的孔雀草种质遗传多样性分析

为了了解孔雀草种质间的亲缘关系,提高孔雀草种质的利用效率,从18个形态特征对40份孔雀草种质进行了遗传变异分析、主成分分析及聚类分析。结果表明,孔雀草表型多样性丰富,种质间表型性状变异程度高,变异系数(CV值)范围3.92%~46.25%,以花朵数最大,其次是冠幅和株高;Shannon-Weaver多样性指数则以叶片性状及花性状较高,平均2.0以上。国内种质比国外种质多样性更丰富。通过主成分分析,筛选出对总体方差累计贡献率达78.949%的4个主成分,并筛选出综合性状表现良好的种质10份。参试的40份种质在欧氏距离阈值为7.29处可分为两大类,一类植株生长势较强包含2份种质(水星黄色和橙色);其余38份种质为一类,在欧氏距离阈值为6.0处,又以花色差异可进一步划分为2个类群,说明生长势及花色可作为孔雀草种质依据表型性状分类的重要指标。     

Moderately acidophilic mesorhizobia isolated from chickpea

AIMS: Our aim was to evaluate the effect of acid and alkaline pH on chickpea rhizobia, and on chickpea-rhizobia symbiosis. METHODS AND RESULTS: Forty-seven rhizobia isolates obtained from 12 Portuguese soils were grown at pH 5, 7 and 9. Among these, 26 grew more at pH 5 than at 7, suggesting the existence of acidophiles. All isolates were identified as mesorhizobia by 16S rDNA partial sequence analysis. Molecular phylogeny of isolates based on partial 16S rDNA sequences suggests that pH tolerance might be species related. Further studies were conducted with six isolates, which were able to grow at acid pH. Isolates PT-35 and 64b grow optimally at pH 6-5-7, with a minimal pH range from 5 to 3, and may thus be considered as moderately acidophilic. Both isolates belong to a previously identified putative new Mesorhizobium species, based on 16S rDNA sequence. CONCLUSIONS: Two moderately acidophilic mesorhizobia isolated from chickpea were identified (PT-35 and 64b). A positive correlation was found between the symbiotic effectiveness at low pH and the acid tolerance of rhizobia isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on moderately acidophilic mesorhizobia, and is an important contribution for the development of highly effective inoculants for chickpea in acid soils.     

Analysis of Growth and Productivity of Green Chickpea Using Nitrogen and Phosphorus Fertilization

Chickpea contains high levels of protein, vitamins and minerals. Acceptable chickpea yield is the result of meeting nitrogen and phosphorus requirements. The effect of appropriately meeting such requirements reflects on growth and can easily be evaluated using growth analysis. This research determined: (a) The effect of nitrogen and phosphorus fertilization on phenology, net assimilation rate, number of green leaves, leaf area, leaf area index and leaf area duration; (b) Green chickpea yield and number of pods due to fertilization; and (c) The combination of nitrogen and phosphorus fertilization that yields the most net revenue. Nitrogen and phosphorus fertilization was evaluated; each at the doses of 0, 75 and 150 kg ha^–1 (N0, N75, N150; P0, P75 and P150, respectively). The combination of the levels of both nutrients generated nine combinations of treatments which were distributed in the field in a randomized complete block design in an arrangement of divided plots with four repetitions. Timing of phenological phases were similar among treatments. Nitrogen and phosphorus fertilization increased number of leaves, leaf area index, and leaf area duration that translated into increased green chickpea yield (GCY). Combinations N150-P75 and N150-P150 produced the highest GCY. The highest net revenue and revenue per peso invested was obtained with N150-P75.     

Studies on the expression of marker genes in chickpea

Conditions were established for the optimum transient expression of beta-glucuronidase and neomycin phosphotransferase II genes introduced into zygotic embryos of chickpea (Cicer arietinum L. 6153 and CM72) by accelerated tungsten particles. Plasmid DNA at a concentration of 12 microgram per milligram of tungsten particles when accelerated with an inflow of helium gas at 60 kilogram per square centimeter through a distance of 24 centimeter in a chamber maintained at a negative pressure of 71.12 centimeter of mercury, resulted in optimal transient expression of the beta-glucuronidase gene in chickpea embryos. However, the expression of the marker genes was 20-40% higher under a cauliflower mosaic virus promoter in comparison to the Win6 and actin promoters. When Agrobacterium tumefaciens was used to transfer marker genes into zygotic embryos and the resultant plants were analysed for activity of the beta-glucuronidase and neomycin phosphotransferase II genes, both of these genes were expressed in tumorous tissues. When a disarmed strain of Agrobacterium was used, normal shoots were regenerated in which the lower parts showed expression of both genes at a frequency of 20%. This revised version was published online in June 2006 with corrections to the Cover Date.     

木豆种质资源形态与农艺性状的多样性分析

为挖掘优异木豆(Cajanus cajan)种质资源,对10份木豆种质资源的10个质量性状和18个数量性状的遗传多样性进行了研究,并对其农艺性状进行了聚类分析。结果表明,质量性状的遗传多样性指数均较大,以鲜荚色(1.9219)的最高,其次是旗瓣点缀色、鲜籽粒颜色和干籽粒底色,多样性指数均为1.4855;再次为干籽粒色斑、干籽粒脐环色和有无种阜,均为0.8813;最小的是小叶叶形、旗瓣底色和株型,均为0.7219。聚类分析可将10份木豆种质资源划分为中茎稀疏型、中茎密生型和粗茎密生型3大类型。这为木豆品种选育提供了科学依据。     

云南薏苡种质资源农艺性状的主成分和聚类分析

为了揭示云南薏苡种质资源多样性,发掘薏苡资源中的有益基因,利用主成分分析和聚类分析,对收集的65个薏苡种质资源的13个农艺性状进行多样性评价。结果表明,云南薏苡资源存在丰富的遗传多样性,其中栽培种的分枝数和分蘖数的遗传变异系数分别达到57.4%和47.5%,野生种百粒重的遗传变异系数达到60.4%。应用主成分分析将云南薏苡13个性状简化为7个主成分,其累积贡献率为85.67%,以叶片宽因子贡献率最高,为49%。采用系统聚类分析,将65份供试材料在遗传距离16.21水平上聚为5个大类,可区分为株高较矮叶片较短型、株高较高叶片较长型以及3个特殊型。     

Strains of Mesorhizobium amorphae and Mesorhizobium tianshanense, carrying symbiotic genes of common chickpea endosymbiotic species, constitute a novel biovar (ciceri) capable of nodulating Cicer arietinum

AIMS: To identify several strains of Mesorhizobium amorphae and Mesorhizobium tianshanense nodulating Cicer arietinum in Spain and Portugal, and to study the symbiotic genes carried by these strains. METHODS AND RESULTS: The sequences of 16S-23S intergenic spacer (ITS), 16S rRNA gene and symbiotic genes nodC and nifH were analysed. According to their 16S rRNA gene and ITS sequences, the strains from this study were identified as M. amorphae and M. tianshanense. The type strains of these species were isolated in China from Glycyrrhiza pallidiflora and Amorpha fruticosa nodules, respectively, and are not capable of nodulating chickpea. These strains carry symbiotic genes, phylogenetically divergent from those of the chickpea isolates, whose nodC and nifH genes showed more than 99% similarity with respect to those from Mesorhizobium ciceri and Mesorhizobium mediterraneum, the two common chickpea nodulating species in Spain and Portugal. CONCLUSIONS: The results from this study showed that different symbiotic genes have been acquired by strains from the same species during their coevolution with different legumes in distinct geographical locations. SIGNIFICANCE AND IMPACT OF THE STUDY: A new infrasubspecific division named biovar ciceri is proposed within M. amorphae and M. tianshanense to include the strains able to effectively nodulate Cicer arietinum.     

271份豌豆种质资源农艺性状遗传多样性分析

以来自五大洲57个国家和地区的271份豌豆资源为材料,利用主成分分析和聚类分析的方法,评价其2个质量性状和7个数量性状的遗传变异水平。结果表明,参试资源的农艺性状具有丰富的遗传多样性。其中遗传多样性指数最高的是始荚节位(2.0590),其次是主茎节数(2.0421);性状变异系数最大的是小区产量(64.874%),其次是百粒重(61.870%)。采用SPSS 22.0软件对参试资源7个数量性状的主成分分析结果表明,前3个主成分因子累计贡献率达66.021%,Ward法聚类将参试的271份豌豆资源划分为4大类群,其中第Ⅱ类群属于高秆、大粒、高产种质,具有很高的丰产潜力,为我国杂交育种亲本选择提供了原材料。对取材数大于等于5的来自于四大洲17个国家的参试资源进一步分析发现,不同地理区域资源间遗传变异显著,其中印度的遗传多样性指数最高(1.7533),巴基斯坦的最低(0.9685),波兰的变异系数和遗传多样性都较高。综合分析国外种质资源的农艺性状,有助于我国豌豆育种项目亲本选配。     

谷子抽穗期与农艺性状的相关与回归分析

抽穗期是农作物的重要性状,决定着作物的地区和季节适应性。明确不同谷子品种适合的种植区域,对生产实践具有重要意义。在海南、洛阳、吉林3个地区连续2年调查了160份谷子资源的抽穗期、株高、穗长、穗重等9个主要性状。相关性分析发现3个地区抽穗期与千粒重均呈负相关,而在海南、洛阳2个地区抽穗期与株高、叶片数、穗长、穗粗、穗重、穗码数、穗粒重均呈正相关,吉林地区抽穗期与穗粒重呈负相关,说明随着抽穗期的适当延长,谷子主要通过增加子粒数目提高产量,而抽穗期过度推迟,则可能导致产量潜力下降;方差分析表明,3个地区抽穗期对千粒重、株高、叶片数、穗长、穗粗、穗重、穗粒重、穗码数均有显著影响(P<0.05)。在海南,随着抽穗期的延长穗粒重递增,抽穗期40 d以上品种穗粒重最大;在洛阳,抽穗期50～60 d的品种穗粒重最高;在吉林,抽穗期70～80 d的品种穗粒重最高。求得每个品种在3个地区的穗粒重均值,以此为依据筛选出53份广生态适应性品种,在3个地区成功建立了叶片数、株高、穗码数对抽穗期的最佳回归方程。本研究表明一定抽穗期范围内谷子主要通过增加穗粒数而不是千粒重来实现增产,筛选出的广生态适应性谷子资源以及建立的回归方程为开展广适应育种、利用抽穗期信息对叶片数、株高、穗码数进行准确选择奠定了基础。     

Effects of nitrogen supply and high temperature on the growth and physiology of the chickpea

Chickpeas were grown with or without nitrate nitrogen feeding, or nodulated with Rhizobium leguminosarum. High [40°C day, 25°C night (HT)] and moderate [25°C day, 177°C night (LT)] temperature regimes were employed during growth. Growth rates, photosynthetic capacity and enzymes of carbon and nitrogen metabolism were monitored to assess the acclimatory capacity of the chickpea. Initial growth rates were stimulated by high temperatures, particularly in nitrate-fed and nodulated plants. Older HT plants had fewer laterals, smaller leaves, and fewer flowers were produced than in LT plants. There was some indication of an acclimation of photosynthesis to high temperatures and this was independent of nitrogen supply. Rubisco activity was increased by high growth temperatures. However, HT plants also had higher transpiration rates and lower water use efficiency than LT plants both in respective growth conditions and when compared in a common condition. High temperatures reduced shoot nitrate reductase activity but had little effect on root activity, which was the same if not greater than activity in LT roots. The amino acid, asparagine, was found at high concentrations in all treatments. Concentrations were maintained throughout growth in HT plants but declined with age in LT plants.     

番茄ILs果实性状的主成分分析与聚类分析

利用以栽培番茄Lycopersicon esculentum(加工番茄M82)为背景创建的L.pennellii LA716渐渗系群体(ILs,introgression lines),对7个番茄果实主要性状进行了主成分和聚类分析。结果表明,7个果实性状可简化为3个主成分,分别为果实质量因子、果形因子和品质因子,累计贡献率85.435%。利用欧式距离,类平均法可将77份渐渗系分为3大类群,第Ⅰ类群包括70个渐渗系材料,在D=17.53的水平又可将第Ⅰ类群分为2个亚群,果实性状较好的材料主要集中在这个类群中;第Ⅱ类群包括1个材料,说明此材料的独特性;第Ⅲ类群包括6个材料。     

Active extrusion of protons and exudation of carboxylic acids in response to iron deficiency by roots of chickpea (Cicer arietinum L.)

A chickpea cultivar, K-850, acidified the nutrient solution in response to iron deficiency, with subsequent re-greening of chlorotic leaves. No recovery of chlorosis was observed when the nutrient solution was buffered at a pH 6.3. During the period of acidification induced by iron deficiency, the roots of K-850 exuded more carboxylic acids than when supplied with sufficient iron. However, the rate of extrusion of protons was much higher than the rate of exudation of carboxylic acids during the acidification period. The extrusion of protons was inhibited by the addition of vanadate at the beginning of the decrease in pH. It appeared that acidification of the solution in response to iron deficiency was mediated by a proton-pumping ATPase, located at the plasma membrane. The presence of cations in the solution was essential for the extrusion of protons under iron deficiency, but the species of cation made no significant difference to the rate of extrusion of protons from roots. Therefore, we concluded that non-specific H+/cation antiport was involved in the acidification process.