Assessment of genetic diversity in clover species from Sardinia, Italy, using AFLP analysis

Two species, Trifolium glomeratum and T. nigrescens, from Sardinia, Italy, were analysed for genetic diversity using amplified fragment length polymorphism (AFLP). Variation between and within populations was compared between the inbreeder, T. glomeratum, and the outbreeder, T. nigrescens. Four AFLP primer combinations resulted in a total of 292 loci, of which 75% were polymorphic in T. glomeratum and 85% in Trifolium nigrescens. Variation was highest between populations in both species, but the difference between populations was greater in T. glomeratum (Fst = 0.17), compared with T. nigrescens (Fst = 0.02). Cluster analysis and principal coordinates analysis were used to verify the relationships found. The high level of genetic variation within populations in both species is attributed to the movement of sheep between paddocks, the existence of both species in Sardinia for thousands of years and the persistence of a long‐lived seedbank due to the production of large numbers of small seeds with high levels of hard seededness.     

Genetic Variations and Adaptations in Chickpea

A total of 329 chickpea lines, comprising 130 kabiili and 199 desi types, originating from six regions, viz., Indian sub-continent, Middle East, North Africa, America, Europe and USSR, were sown in augmented design. The data recorded on agronomic characters on all entries from each region were evaluated in terms of group (region) means. Among the six regions, greater plant height and lateness in flowering were specific to USSR entries. Group means of USSR accessions, both desi and kabuli, differed significantly from those of Indian varieties for a majority of the characters. These observations indicate that the degree of expression of certain characters could be ascribed to specific areas, which led to area-specific adaptations. Furthermore, type-specific differentiating characters, differing in degree of expression, were observed in desi and kabuli types. For example, high mean values of characters, like branches, pods, grain yield and harvest index, were associated with desi types, and 100-seed weight with kabulis. History of cultivation and selection for specific purpose has been cited as cause for area-specific adaptations and type-specific differentiating characters.     

Exploitation of wild annual Cicer species for widening the gene pool of chickpea cultivars

Introgression of unadapted genes from the wild Cicer species could contribute to the widening of genetic base of important traits such as yield, yield attributes and resistance to major biotic and abiotic stresses. An attempt was made successfully to intercross two wild annual Cicer species with three cultivated chickpea cultivars. Four interspecific cross‐combinations were made, and their true hybridity was ascertained through morphological and molecular markers. These cross‐combinations were also studied for some important quantitative traits under real field conditions. The range, mean and coefficient of variation of agro‐morphological traits were assessed in the parental lines, their F₁ and F₂ generations to determine the extent of variability generated in cultivated chickpea varieties. A high level of heterosis was recorded for number of pods/plant and seed yield/plant in F₁ generation. Three cross‐combinations of ‘Pusa 1103’ × ILWC 46, ‘Pusa 256’ × ILWC 46 and ‘Pusa 256’ × ILWC 239 exhibited substantially higher variability for important yield‐related traits. The present research findings indicate that these wild annual Cicer species can be easily exploited to broaden the genetic base of cultivated gene pool for improving seed yield as well as adaptation.     

Genotypic Variation in Root Systems of Chickpea (Cicer arietinum L.) across Environments

Root systems of various chickpea genotypes were studied over time and in diverse environments, – varying in soil bulk density, phosphorus (P) levels and moisture regimes. In a pot study comparing a range of chickpea genotypes, ICC 4958 and ICCV 94916‐4 produced higher root length density (RLD) and root dry weight (RDW), which were better expressed under P stress conditions. In two field experiments in soils of intermediate and high soil bulk densities, ICC 4958 also had greater RLD and RDW, particularly under soil moisture stress conditions. The expression of greater rooting ability of ICC 4958 under a wide range of environmental conditions confirms its suitability as a parent for genetically enhancing drought resistance and P acquisition ability. The superiority of ICC 4958 over other genotypes was for root proliferation expressed through RLD. Thus, the variation in RLD can be the most relevant root trait that reflects chickpea's potential for soil moisture or P acquisition.     

Genetic diversity in cornsalad (Valerianella locusta) and related species as determined by AFLP markers

Fifteen amplified fragment length polymorphism (AFLP) EcoRI/MseI‐based primer combinations with five selective bases (Eco RI‐ANN, MseI‐CN) were used to estimate genetic diversity among 45 line varieties of cultivated cornsalad and 19 genebank accessions classified into nine different species related to cornsalad. Polymorphic fragments were scored for calculation of Jaccard's coefficient of genetic similarity (GS). The average GS estimate in elite germplasm (GS = 0.90) was substantially higher than in exotic germplasm (GS = 0.47). UPGMA‐cluster analysis revealed genetic relationships among recently bred varieties, old varieties and genebank accessions. Analysis of molecular variance indicated almost threefold variability within sets compared with between sets due to a high level of polymorphism among wild species. Sources for increasing genetic diversity in elite germplasm of cornsalad were suggested and a duplicate among the genebank accessions was detected. AFLPs could be considered a powerful tool for genetic diversity estimation in cornsalad germplasm and are recommended for systematic fingerprinting of remaining cornsalad species.     

Intraspecific olive diversity assessed with AFLP

Amplified fragment length polymorphism (AFLP) was used to study diversity within and among Spanish olive varieties. A high degree of diversity was found among the varieties present in each growing region. Olive oil production and quality relies on appropriate cultivar selection as well as good orchard management. Production based only on a few superior cultivars would result in improved yield, oil quality, and production management. Amplified fragment length polymorphism were evaluated as a tool to identify the intraspecific and intravarietal diversity of olive. Amplified fragment length polymorphism analysis of 38 accessions belonging to 10 cultivars using six primer combinations produced 106 polymorphic bands. Results were analyzed for similarity among accessions via unweighted pair‐group means cluster analysis, resulting in 10 clusters corresponding to named variety designations. Similarity among varieties ranged from 0.60 to 0.72. Diversity within varieties was identified. Similarity within named varieties (accessions with the same varietal name) ranged from 0.75 to 0.96. Differences in several markers were found among 34 accessions. Intravarietal diversity was shown to exist within the Spanish olive varieties grown in the region surrounding Valencia.     

Pod volume and pod-filling percentage as additional traits for the characterization of chickpea (Cicer arietinum L.)

Thirty short- to medium-duration chickpea germplasm accessions from diverse geographic origins and with a wide range of physiological and morphological traits were grown in three environments at ICRISAT Asia Center, Patancheru, during 1992/93. Data were recorded on time to flowering, leaf area, 100-seed mass, pod volume, and pod-filling percentage. Quantitative data on the last two traits were recorded for the first time to examine their relevance to the characterization of germplasm accessions and their use as selection criteria in breeding. The accessions exhibited considerable variation for the traits. The broad-sense heritabilities were 0.98 for pod volume and 0.85 for pod-filling percentage. The two traits showed consistent relationships with other morphological characters indicating that the pod volume and pod-filling percentage traits can be utilized in genotype characterization of chickpea.     

Genetic control of seed weight and calcium concentration in chickpea seed

Chickpea, Cicer arietinum L., is a staple protein source in many Asian and Middle Eastern countries. Hence, the mineral content of its seed, especially that of calcium, is of nutritional importance. Calcium is transported through plants and to legume pods almost exclusively via the xylem stream, with Ca accretion in developing seeds resulting primarily from diffusion of Ca from the adjoining pod wall. Thus, for seeds of differing surface‐to‐mass ratios, Ca concentration is expected to correlate inversely with seed weight. The relationship between seed weight and Ca concentration in chickpea seeds was studied using a range of germplasm and derivatives from crosses between types differing in seed Ca concentration. Among the cultivars tested, low seed mass was associated with high Ca concentration. However, the study of hybrid progeny indicated that seed Ca content was mainly determined by genetic factors other than grain weight genes. This finding may assist in future breeding of high nutritional quality chickpea cultivars.     

Inheritance of Dry Root Rot (Rhizoctonia bataticola) Resistance in Chickpea (Cicer arietinum)

The inheritance of resistance to dry root rot of chickpea caused by Rhizoctonia bataticola was studied. Parental F₁ and F₂ populations of two resistant and two susceptible parents, along with 49 F₁ progenies of one of the resistant × susceptible crosses were rested for their reaction to dry root rot using the blotting-paper technique. All F, plants of the resistant × susceptible crosses were resistant; the F₂ generation fitted a 3 resistant: 1 susceptible ratio indicating monogenic inheritance, with resistance dominant over susceptibility. F³ family segregation data confirmed the results. No segregation occurred among the progeny of resistant × resistant and susceptible × susceptible crosses.     

AFLP in Triticum aestivum L.: patterns of genetic diversity and genome distribution

The amplified fragment length polymorphism (AFLP) procedure was applied to a diverse panel of wheat (Triticum aestivum L. em. Thell.) accessions and sixty-nine of the recombinant inbred lines (RILs) from the widely used genetic mapping population derived from the cross of Opata 85 and W7984. Most (76.8%) bands were monomorphic among T. aestivum accessions. The majority of bands monomorphic in T. aestivum also were present in the synthetic wheat parent (W7984). Ten primer pairs generated 153 polymorphic AFLP bands, 140 of which could be assigned to a chromosome location and were relatively evenly distributed on the genetic linkage map. AFLP loci in T. aestivum were distributed throughout the genome; they generally have only one detectable sequence variant; and they exhibit monogenic dominant mendelian inheritance. Frequencies of polymorphic bands in the germplasm sampled are in the range that enables informative cluster analyses as well as map-based diversity and association analysis studies. AFLP bands mapped to individual loci in the Opata 85/W7984 RIL population will frequently be polymorphic in other crosses or germplasm, irrespective of whether the band arises from the T. aestivum parent or the synthetic wheat parent. This revised version was published online in July 2006 with corrections to the Cover Date.     

Relationship Between Cold Severity and Yield Loss in Chickpea (Cicer arietinum L.)*

Seed yield in chickpea (Cicer arietinum L.) is substantially increased by advancing sowing date from the traditional spring to early winter at low to medium elevation areas around the Mediterranean Sea. This shift, however, increases the probability of the exposure to subzero temperatures as low as -10 °C for up to 60 days in a year. These low temperatures often reduce seed yield of cold-susceptible cultivars. Yield losses from cold were estimated in two experiments conducted at Tel Hadya, Syria. In experiment 1, of 96 genotypes sown on nine dates ranging from autumn to spring during the 1981–82 season, those lacking tolerance to cold were killed and produced no yield in autumn sowing, whereas lines with cold tolerance produced nearly 4 t/ha which corresponds to a four-fold increase over spring sowing. Moderately cold-tolerant genotypes sown during early winter produced substantially more seed yield than the normal spring-sown crop. Seedlings were more cold tolerant than the plants in early or late vegetative stages. In experiment 2, in which yield loss due to cold in the field was estimated in 12 yield trials comprising 288 newly bred lines in the 1989–90 season, the regression of cold susceptibility on seed yield in each of the trials was highly significant and negative. On average, winter-sown trials produced 67 % more seed yield than spring-sown trials, but 125 out of 288 genotypes produced yield more than double in winter sowing. Early maturing lines suffered severe cold damage and many lines produced no seed.     

Inheritance of Seed Coat Thickness in Chickpea (Cicer arietinum L.) and its Evolutionary Implications

The desi and kabuli chickpeas are characterized, among other things, by their seed coats being thicker in the desi than in the kabuli type. The inheritance of seed coat thickness, and its relation to flower colour and seed size, was studied. Seed coat thickness exhibits monogenic inheritance, the thin kabuli seed coat being the recessive character. Linkage was found between seed coat thickness and flower colour, the recombinant fraction being 0.19. No relationship was found between seed coat thickness and seed size. The role of these characters in the evolution of the chickpea is discussed.     

Interspecific hybridization between Cicer arietinum and C. pinnatifidum

An interspecific hybrid between Cicer arietinum cv. GL 769 and a wild species C. pinnatifidum was obtained after emasculation, pollination and application of growth regulators. Ovules were cultured and embryos were later dissected to obtain hybrid plants. These plants were albinos and morphologically resembled C. pinnatifidum. Shrivelled seeds were also obtained in 2% of the crosses, which on germination gave rise to albino plants. These plants did not survive beyond 20 days. The hybrid nature of these plants was confirmed by esterase isozyme studies. Hybrid shoots obtained from germinating embryos were cultured on modified ML-6 medium with BAP 2 mg/1, IAA 0.5 mg/1, where they turned green after 3–4 weeks. Transmission electron microscopy (TEM) studies on leaf sections from green hybrid shoots showed an improvement in the chloroplast structure, with better organized grana.     

Two new traits - open flower and small leaf in chickpea (Cicer arietinum L.)

Two new traits – open flower and small leaf in chickpea are discussed. Open flower, a natural mutant in a good agronomic background is reported for the first time, small leaf trait has been reported earlier, and has now been studied by breeders. Both useful traits were found to be monogenic recessive. The joint F₂ segregation data revealed no linkage between flower colour and flower type, but flower type and leaf size showed some linkage. Open flower could contribute to a higher rate of cross pollination and utilization of heterosis. The small leaf allows light to penetrate the crop canopy, and could be useful in designing a physiologically efficient plant type in chickpea. This revised version was published online in July 2006 with corrections to the Cover Date.     

人工合成甘蓝型油菜早期世代基因组变异的AFLP和MSAP标记研究

为了揭示人工甘蓝型油菜早期世代遗传和表观遗传变异规律, 以A组合(大黄油菜×中花芥蓝) S₀世代、B组合(大黄油菜×中迟芥蓝) S₀和S₁世代人工甘蓝型油菜为材料, 分别利用AFLP和MSAP技术检测基因组变化及甲基化模式变化情况。结果表明, 16对引物在A组合S₀扩增到523条带, 其中4对引物扩增出9条变异带, 包括7条亲本缺失带和2条新增带, 分别占S₀总条带的1.33%和0.38%;45对引物在B组合双亲植株扩增到1093条带, 只有1对引物检测到1条父本带型在所有S₀植株中缺失, 约占S₀总条带的0.09%;在B9子代F19-1~F19-16总共扩增得到1092条带, 变异带有10条, 占总条带的0.915%, 其中包括9条缺失带和1条新增带, 9条缺失带全部位于C基因组。MSAP检测发现, B组合S₀植株中有3个位点发生了甲基化模式的改变, 全部位于A基因组, 甲基化模式改变位点占总检测位点的1.37%。研究还发现B组合S₀世代一个植株出现可遗传的花色变异, 推测该表型变异与B组合人工甘蓝型油菜中C基因组变异有关。     

梨种质资源遗传多样性研究中的AFLP技术引物筛选

本研究对带有3个选择性核苷酸的EcoRI和MseI引物各8个所组成的64个引物组合进行筛选,选出了多态性高的引物10对用于正式扩增。所选出的引物组合共扩增出371个位点,其中多态性位点306个,占总位点数的82.5%,每个引物组合扩增的位点数变化在23-55个之间,平均37.1个。所筛选出的10个引物组合为梨属植物AFLP扩增多态性引物,可广泛用于梨属植物的AFLP扩增技术。中国梨属植物资源丰富,通过对AFLP多态性引物的筛选,为该技术在梨属种质资源中的应用提供参考。