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1.
The selection of Jatropha based on morphological information and molecular markers is essential as it is more reliable and consistent. Hence, twelve Jatropha accessions from different geographical areas of India were screened for genetic diversity using 19 morphological traits and 21 ISSR primers. The analysis of morphological traits grouped the accessions into five clusters. The cluster I consisted of J. curcas (CJC 18), J. curcas (CJC 20), J. curcas (CJC 22), J. curcas (CJC21), and J. curcas (CJC 25), and contained the maximum number of accessions; clusters II and IV contained the minimum number of accessions. Among all the characters, the highest range was exhibited by plant height and the least value by the number of branches. The twenty-one ISSR primers generated 156 polymorphic alleles. The average number of ISSR alleles generated was 7.47 per primer. The ISSR primer UBC 884 was highly informative with the maximum of 12 alleles. The 12 genotypes were grouped into eight clusters. The cluster I contained the maximum number of accessions, namely J. curcas (CJC 18), J. curcas (CJC 20), J. curcas (CJC 22), J. curcas (CJC21), and J. curcas (CJC 25). The clusters II, III, IV, V, VI, VII, and VIII (J. tanjorensis, J. gossypiifolia, J. glandulifera, J. podagrica, J. ramanadensis J. villosa, and J. integerrima) contained the minimum number of accessions. Maximum diversity between J. villosa and J. integerrima was noticed and the least diversity between J. curcas (CJC21) and J. curcas (CJC 25) seen because the ISSR markers differentiated the Jatropha accession into a wide genetic diversity as compared to the morphological data. The species-specific diagnostic markers identified in the study such as 1000 bp alleles for J. glandulifera by the primer UBC 826 is suitable for discriminating species of Jatropha, and thus can be used for identifying a Jatropha species from any mixed population comprising other members of the Jatropha complex.  相似文献   

2.
This paper describes the relative efficiency of three marker systems, RAPD, ISSR, and AFLP, in terms of fingerprinting 14 rice genotypes consisting of seven temperatejaponica rice cultivars, three indica near-isogenic lines, three indica introgression lines, and one breeding line of japonica type adapted to high-altitude areas of the tropics with cold tolerance genes. Fourteen RAPD, 21 ISSR, and 8 AFLP primers could produce 970 loci, with the highest average number of loci (92.5) generated by AFLP. Although polymorphic bands in the genotypes were detected by all marker assays, the AFLP assay discriminated the genotypes effectively with a robust discriminating power (0.99), followed by ISSR (0.76) and RAPD (0.61). While significant polymorphism was detected among the genotypes of japonica and indica through analysis of molecular variance (AMOVA), relatively low polymorphism was detected within the genotypes of japonica rice cultivars. The correlation coefficients of similarity were significant for the three marker systems used, but only the AFLP assay effectively differentiated all tested rice lines. Fingerprinting of backcross-derived resistant progenies using ISSR and AFLP markers easily detected progenies having a maximum rate of recovery for the recurrent parent genome and suggested that our fingerprinting approach adopting the ‘undefined-element-amplifying’ DNA marker system is suitable for incorporating useful alleles from the indica donor genome into the genome of temperate japonica rice cultivars with the least impact of deleterious linkage drag.  相似文献   

3.
There is a recent surge in the marker‐assisted selection of desired sex type among economically important dioecious crops. Simmondsia chinensis (Jojoba), a dioecious crop is of immense agricultural importance where only the female plants are preferred for commerce. DNA fingerprinting technology, ISSR analysis along with bulk segregant analysis (BSA), has been carried out on a diverse set of 17‐ to18‐year‐old mature male and female plants of Jojoba to validate a male sex‐specific ISSR marker, UBC‐8071200 on larger population that comprises 330 female and 255 male plants of Jojoba. This male sex‐specific DNA fragment of ~1200 bp was cloned and sequenced. The sequence was found to be 1120 bp in length and based on the sequence information, a pair of sequence tagged sites primers was developed that amplified a single ~800 bp band, consistently only in all the male populations while no amplification was seen in their female counterparts. The marker was named as Jojoba Male Sex Marker which was further validated on 330 female plants from 22 genotypes and 255 male plants from 17 genotypes.  相似文献   

4.
Molecular markers provide novel tools to differentiate between the various grades of Basmati rice, maintain fair-trade practices and to determine its relationship with other rice groups in Oryza sativa. We have evaluated the genetic diversity and patterns of relationships among the 18 rice genotypes representative of the traditional Basmati, cross-bred Basmati and non-Basmati (indica and japonica) rice varieties using AFLP, ISSR and SSR markers. All the three marker systems generated higher levels of polymorphism and could distinguish between all the 18 rice cultivars. The minimum number of assay-units per system needed to distinguish between all the cultivars was one for AFLP, two for ISSR and five for SSR. A total of 171 (110 polymorphic), 240 (188 polymorphic) and 160 (159 polymorphic) bands were detected using five primer combinations of AFLP, 25 UBC ISSR primers and 30 well distributed, mapped SSR markers, respectively. The salient features of AFLP, ISSR and SSR marker data analyzed using clustering algorithms, principal component analysis, Mantel test and AMOVA analysis are as given below: (i) the two traditional Basmati rice varieties were genetically distinct from indica and japonica rice varieties and invariably formed a separate cluster, (ii) the six Basmati varieties developed from various indica × Basmati rice crosses and backcrosses were grouped variably depending upon the marker system employed; CSR30 and Super being more closer to traditional Basmati followed by HKR228, Kasturi, Pusa Basmati 1 and Sabarmati, (iii) AFLP, ISSR and SSR marker data-sets showed moderate levels of positive correlation (Mantel test, r = 0.42–0.50), and (iv) the partitioning of the variance among and within rice groups (traditional Basmati, cross-bred Basmati, indica and japonica) using AMOVA showed greater variation among than within groups using SSR data-set, while reverse was true for both ISSR and AFLP data-sets. The study emphasizes the need for using a combination of different marker systems for a comprehensive genetic analysis of Basmati rice germplasm. The high-level polymorphism generated by SSR, ISSR and AFLP assays described in this study shall provide novel markers to differentiate between traditional Basmati rice supplies from cheaper cross-bred Basmati and long-grain non-Basmati varieties at commercial level.The first two authors have equal contribution  相似文献   

5.
为创建砂仁及其主要混伪品益智仁的ISSR分子鉴别方法,以UBC818为引物,对影响ISSR-PCR反应体系的引物、dNTPs、DNA模板、Taq DNA聚合酶和Mg2+浓度进行5因素4水平正交优化试验,并在此基础上筛选阳春砂ISSR引物及砂仁正伪品的鉴别引物.结果表明,20 μL阳春砂ISSR-PCR最佳反应体系包括引...  相似文献   

6.
A recombinant inbred line (RIL) mapping population (F8) was generated by crossing Vigna mungo (cv. TU 94‐2) with Vigna mungo var. silvestris and screened for mungbean yellow mosaic virus (MYMV) resistance. The inter simple sequence repeat (ISSR) marker technique was employed to identify markers linked to the MYMV resistance gene. Of the 100 primers screened, 54 showed amplification of which 36 exhibited polymorphism between the parents TU 94‐2 (resistant) and V. mungo var. silvestris (susceptible). Individual plants from 53 RIL populations were analysed and one marker (ISSR8111357) was identified as tightly linked to the MYMV resistant gene at 6.8 cM. Both the phenotype as well as the ISSR8111357 marker segregated in a 1 : 1 ratio. The ISSR8111357 marker was sequenced and sequence characterized amplified region (SCAR) primers were designed (YMV1‐F and YMV1‐R) to amplify the marker. Screening for the SCAR marker in the RIL population distinguished the MYMV resistant and susceptible plants, agreeing well with the phenotypic data. The ISSR8111357 marker was validated using diverse blackgram genotypes differing in their MYMV reaction. The marker will be useful for the development of MYMV‐resistant genotypes in blackgram.  相似文献   

7.
K. Vijayan    P. P. Srivatsava    C. V. Nair    A. K. Awasthi    A. Tikader    B. Sreenivasa    S. R. Urs 《Plant Breeding》2006,125(3):298-301
Mulberry (Morus indica L.) is an important tree crop being exploited for feeding the silk‐producing insect Bombyx mori L. In order to identify parents suitable for breeding to raise high‐yielding varieties for the non‐traditional areas of Kerala, India and also to identify markers associated with leaf yield attributing traits, the present study was undertaken with 44 mulberry genotypes. Variability on morpho‐biometric traits and molecular markers, generated with 12 selected ISSR primers, was estimated. Significant differences between genotypes were observed for all the traits. The dendrogram generated with morpho‐biometric characters clustered the genotypes into three distinct groups and one isolate, while the same using Inter simple sequence repeat (ISSR) markers clustered the genotypes into five groups and six isolates. The greater resolving power of the ISSR markers was evident form the dendrograms. Using step‐wise multiple regression analysis, a number of markers associated with number of branches, total shoot length, leaf weight, internodal distance, leaf chlorophyll, protein, leaf moisture percentage were identified. These markers could be of much use in Marker assisted selection (MAS) breeding programmes in mulberry, especially when no genetic information in terms of linkage maps and Quantitative Trait Locis (QTLs) is available a plant with high heterozygosity and a long juvenile period.  相似文献   

8.
本研究采用均匀设计和单因素试验相结合的方法,探寻淫羊藿ISSR-PCR的各组分(即引物, 2×Taq Master Mix,模板DNA)的最佳用量及退火温度对ISSR-PCR扩增的影响,为进一步使用ISSR分子标记分析淫羊藿的遗传多样性提供科学依据。结果表明,筛选的最佳体系为:在20μL的体系中,模板DNA的量为30ng,2×Taq Master Mix的量为9.8μL,引物为0.325μmol/L。此外,筛选出7条多态性较好、条带稳定的引物(UBC-808, UBC814, UBC826, UBC827, UBC840, UBC846及UBC856),并对其进行温度梯度PCR,结果表明,所选引物的最佳退火温度介于46.8℃~65℃之间。在此基础上,对13份淫羊藿种质资源进行ISSR扩增验证,结果表明建立的最佳反应体系扩增效果较好,稳定性强,对淫羊藿的遗传多样性分析、鉴定等具有较好的应用价值。  相似文献   

9.
F. Javidfar    V. L. Ripley    V. Roslinsky    H. Zeinali    C. Abdmishani 《Plant Breeding》2006,125(1):65-71
The quality of the oil derived from oilseed rape is determined by its fatty acid composition. Breeding oilseed rape for enhanced oil quality includes the development of cultivars with high oleic and low linolenic acid. Random amplified polymorphic DNA (RAPD) and intersimple sequence repeat (ISSR) techniques were investigated for the development of molecular markers for genes controlling oleic and/or linolenic acid. Markers that were identified were converted to sequence characterized amplified region (SCAR) markers for use in breeding. Molecular markers associated with these two fatty acids were identified in a doubled haploid population derived from a cross between the oilseed rape lines TO99‐5318‐20, very high oleic (>79%) and very low linolenic acid (<2%) × DH12075, high oleic (68%) and higher linolenic acid (>7%). Eight RAPD markers were associated with oleic and linolenic acid contents. The RAPD marker UBC 2830 accounted for 43% and 13% of the genetic variation for oleic and linolenic acid levels, respectively. The RAPD marker UBC 153550 accounted for 19% of the genetic variation for linolenic acid. The UBC 2830 fragment was converted to a SCAR marker. The markers identified in this study should be useful tools for the early generation selection of high oleic and low linolenic acid genotypes in oilseed rape breeding programmes.  相似文献   

10.
40个龙眼品种(品系)DNA指纹图谱构建及遗传关系分析   总被引:2,自引:1,他引:1  
有效鉴别龙眼新品种(品系),为龙眼种质资源研究和新品种保护提供理论基础。利用ISSR分子标记分析‘冬宝9 号’、‘高宝’等4 个龙眼新品种(品系)与36 个龙眼品种的遗传关系,并建立其DNA指纹图谱。从80 个ISSR 引物中筛选出11 个多态性好、扩增清晰的引物。结果显示,11 个引物共获得146个位点,有125 个多态性位点,多态性百分率为85.7%。40 个龙眼品种(品系)的遗传相似系数在0.58~0.88 之间。UPGMA聚类结果表明,40 个龙眼品种(品系)在遗传相似系数为0.64 的水平处划分为3 组。引物UBC810/UBC818 组合可将40 个龙眼品种(品系)全部区分开,并依此建立了数字化的DNA指纹图谱,为龙眼新品种的鉴别、分类、示范及推广等方面提供依据。  相似文献   

11.
An assessment of genetic diversity studies was undertaken to understand the level and pattern of diversity in 65 mango (Mangifera indica L.) genotypes of India including 20 commercial cultivars, 18 hybrids, 25 local genotypes and two exotic cultivars based on qualitative and quantitative fruit characters as well as RAPD and ISSR profiles. A considerable variation was observed in respect of three important qualitative characters namely table quality, fruit attractiveness and storage life of ripe fruits and potentially superior genotypes for the above traits were identified. A wide variation was noticeable regarding metabolite composition of pulp of ripe mango fruit with respect to total soluble solids, total sugar, reducing sugar, acidity, sugar:acid ratio, ascorbic acid and phenolic content. Fifteen RAPD primers yielded 27 monomorphic and 129 polymorphic bands with percent polymorphism averaging 82.7%. Of a total 70 ISSR bands generated from eight ISSR primers, 60 bands (85.71%) were found to be polymorphic. Cumulative band data from these two methods precisely arranged accessions into eight clusters which correspond well with their pedigree relationship. UPGMA dendrograms drawn using RAPD, ISSR and cumulative data showed highly similar grouping of genotypes on the basis of their parental origin. No clear-cut geographical separation was revealed among East, West, North and South Indian mango cultivars by neither of these molecular markers nor their combinations. This supports the common gene pool origin of mango as well as operation of similar selection pressure as the cultivar preferences in these areas are largely similar.  相似文献   

12.
The genetic variability and relationships among 11 cowpea genotypes representing two cultivars and nine elite genotypes were analyzed using 22 random amplified polymorphic DNA (RAPD) and nine inter-simple sequence repeat (ISSR) markers. ISSR markers were more efficient than RAPD assay with regards to polymorphism detection. But the average numbers of polymorphic loci per primer and resolution power were found to be higher for RAPD than for ISSR. Also, the total number of genotype specific marker loci, Nei’s genetic diversity, Shannon’s information index, total heterozygosity, and average heterozygosity were prominent in RAPD as compared to ISSR markers. The regression test between the two Nei’s genetic diversity indices showed low regression (0.3733) between ISSR and RAPD + ISSR-based similarities but maximum (0.9823) for RAPD and RAPD + ISSR-based similarities. The RAPD- and ISSR-generated cultivar- or genotype-specific unique DNA fingerprints able to identify the most diverse genotypes. A dendrogram constructed based on RAPD and ISSR combined data indicated a very clear pattern of clustering according to the groups (cultivars and elite genotypes). The results of principal coordinate analysis were comparable to the cluster analysis. Cluster analysis showed that most diverse genotypes (GP-125 — small size with good seed quality; GP-129, GP-90L — big size with poor seed quality) were separated from moderately diverse cultivars and genotypes. The genetic closeness among GP-129 and GP-90L, JCPL-42, and JCPL-107 could be explained by the high degree of commonness in these genotypes.  相似文献   

13.
Segregation for salinity tolerance and ISSR markers based molecular polymorphism were investigated in a F3 plant population raised via single-seed descent method from a cross between salt-tolerant indica rice variety CSR10 and salt-susceptible premium traditional Basmati rice variety Taraori Basmati HBC19. A total of 130 F3plants were evaluated individually for salinity tolerance on 1–9 scale on the basis of seedling growth parameters; the average score ranged between 1.7 to 8.3. Frequency distribution curve obtained using the salinity tolerance data of F3 population and a chi-square analysis, showed a good fit to a normal distribution. Eleven plants each in the category of salt-tolerant and salt-susceptible were selected from the segregating F3 population for ISSR marker analysis. A total of 149 bands (4–11 bands per primer) ranging from 200 to 3530 bp were scored for the two rice varieties and the selected CSR10 × HBC19 segregating F3 plants using 26 ISSR primers. Of these, 89 were monomorphic and 60 were polymorphic. Of the 60 polymorphic bands,36 and 20 bands were specific to CSR10 andHBC19 respectively. The remaining four bands were amplified using UBC primers 810,848, 853 and 886 and present in only some of the CSR10 × HBC19 F3 plants. Notably, ISSR primers with dinucleotide repeat motif and 5'-anchored end amplified more number of bands (7.0 bands/primer) compared to3'-anchored dinucleotide primers (5.4bands/primer), but 3'-anchored dinucleotide primers revealed higher level of polymorphism (2.6 polymorphic bands/primer) compared to 5'-anchoreddinucleotide primers (1.43 polymorphic bands/ primer). While distribution of majority of the polymorphic bands were more or less in the expected ratios in salt-tolerant and/or salt-sensitive F3segregating plants, but some of the bands amplified using UBC ISSR primers 823, 825,826, 849, 853, 864, 866 and 884 showed highly skewed distribution. Such polymorphic bands stand greater chances of having a linkage with the genes/ QTLs for salinity tolerance and shall be the target for further studies. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

14.
A collection of 11 coloured cotton Gossypium hirsutum genotypes and four white linted genotypes of different origin were evaluated by randomly amplified polymorphic DNA (RAPD) analysis. These 15 cotton genotypes were evaluated using 32 different 10‐mer primers of arbitrary sequences. All 32 primers were polymorphic ‐ in total, 287 amplified fragments were observed in these patterns. Out of the 287 fragments, 219 were polymorphic accounting for 76.31% of the total number of fragments. Similarity indices were calculated using the Dice coefficient and a dendrogram showing relationships between genotypes was obtained by Unweighted Pair Group Method of Arithmetic Average (UPGMA) cluster analysis. Cluster analysis showed clear‐cut separation of the coloured and white linted genotypes and thus formed three clusters (I, II and III). Among the coloured linted genotypes, all except ‘Parbhani American’ and ‘Lousiana brown’ clustered together. Cluster II contained white linted genotypes orginating from the same breeding station. The results indicate that RAPDs may constitute a relatively simple and efficient method for analysing genetic variation in coloured and white linted G. hirsutum collections.  相似文献   

15.
In this work we detected the extent of variability at nuclear and cytoplasmic DNA level of regenerated plants belonging to Solanum genotypes with a different genetic background and somatic chromosome number. As for the nuclear characterization, a total of 66 (18.5%) polymorphic bands were scored using 13 ISSR primers on 45 randomly selected regenerants. Our results show that the regenerants obtained from clone cmm 1T and, at lower level, those from cph 1C are unstable under in vitro conditions or rather more prone to in vitro-induced stress leading to somaclonal variation than the other genotypes used. Two types of changes were observed: disappearance of parental ISSR fragments, termed “loss”; appearance of novel ISSR fragments, termed “gain”. The most frequent event occurring in the regenerants was the loss of fragments (41 bands). Regenerated plants were analyzed with seven plastid universal primers to determine the cytoplasmic composition at chloroplast level. All cpDNA primer pairs tested produced amplicons of the same size in all genotypes analyzed and no polymorphic fragments were observed with any universal primers used. Our results show that under in vitro culture conditions genotype affects the integrity of the genome. In addition, the absence of polymorphism at plastid level confirms the greater genetic stability of cytoplasmic DNA.  相似文献   

16.
Novel male-specific molecular markers (MADC5, MADC6) in hemp   总被引:8,自引:0,他引:8  
Decamer RAPD primers were tested on dioecious and monoecious hemp cultivars to identify sex-specific molecular markers. Two primers (OPD05 and UBC354) generated specific bands in male plants. These two DNA fragments were isolated, cloned and sequenced. Both markers proved to be unique, since no sequence with significant homology to OPD05961 and UBC354151 markers were found in databases. These markers were named MADC3 (OPD05961) and MADC4 (UBC354151) (Male-Associated DNA from Cannabis sativa). The markers were converted into sequence-characterized amplified region (SCAR) markers. The SCAR markers correlated with the sex of the segregating F2 population and proved the tight linkage to the male phenotype. Results of F2 plant population analysis suggest these markers are to be linked to the Y chromosome. This revised version was published online in August 2006 with corrections to the Cover Date.  相似文献   

17.
Rye (Secale cereale L. and S. strictum) offers potential to increase the genetic variability and to introduce desirable characters for wheat improvements. Cytogenetic techniques have been used to screen wheat lines containing rye chromatin. These techniques are not adequate since they are highly technical and time consuming. They are not suitable for breeding programs that require rapid screening of large numbers of genotypes. The main objective of this study was to develop and characterize ISSR and SCAR markers that can distinguish wheat from rye genome. Total DNA from wheat, rye, and triticale accessions from different provenances were amplified with ISSR primers in PCR assays. Three wheat-diagnostic sequences were identified. In addition three rye-diagnostic ISSR markers of which, one marker specifically diagnostic for Secale strictum were characterized. Pairs of primers flanking these specific sequences were designed to produce SCAR markers. Two SCAR markers were rye genome-specific. One SCAR was present in all the seven rye chromosome, and another was specific to rye chromosomes two, three, four, and seven. These newly developed ISSR and SCAR markers should be useful to wheat breeders screening genotypes that may contain rye chromatins.  相似文献   

18.
19.
The distribution and allelic expressivity of hybrid necrosis genes (Ne 1 and Ne 2) were studied in 21 winter (mostly exotic) and 43 spring type elite wheat genotypes, by crossing them with two known testers, C 306 (Ne 1-carrier) and HD 2380 (Ne 2-carrier).Ne 1 gene was present in one north-west Himalayan winter wheat landrace, Shoure Local, but absent in the other winter as well as spring wheats. Ne 2 gene was prevalent to a much lower extent in the exotic winter wheat germplasm (31.57%) as compared to the recently developed Indian and Mexican spring wheat semidwarfs (69.80%). This may suggest that breeders have tried to preclude hybrid necrosis by selecting for non-carrier genotypes in the development of exotic winter wheats in contrast to the situation in spring wheats. Based on the degree of expression of hybrid necrosis genes in the F1 hybrids, the carrier genotypes were characterized with respect to the allelic strength of the hybrid necrosis genes. The 27 non-carrier genotypes of the two ecotypes identified in the present study have a greater potential use in future hybridization programmes so as to overcome the problem of hybrid necrosis. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

20.
Wild Lens taxa are invaluable sources of useful traits for broadening genetic base of cultivated lentil. Nine inter‐sub‐specific and interspecific crosses were made successfully between cultivated (Lens culinaris ssp. culinaris) and wild lentils (L. culinaris ssp. orientalis, odemensis, lamottei and ervoides). The effect of species groups, day length and temperature on crossability in lentils was evident under normal winter sowing in New Delhi and in summer Himalayan nursery at Sangla in Himachal Pradesh, India, although pollen fertility assessed in all the cross‐combinations showed no significant variation. True hybridity of nine inter‐sub‐specific and interspecific crosses was confirmed through morphological and molecular (ISSR) markers, in which three of 120 primers could confirm the hybridity of all the crosses. All cross‐combinations were also studied for important quantitative traits related to yield. The range, mean and coefficient of variation were estimated in parental lines, F1 and F2 generations to determine the extent of variability generated in cultivated lentils through the introgression of genes from wild L. taxa. A high level of heterosis was observed in F1 crosses for important traits studied. Substantially higher variations for seed yield and its attributing traits were exhibited in F2 generations indicating transgressive segregation. The results of the present investigation revealed that wild L. taxa can be successfully exploited for lentil improvement programmes, and the variations generated could be easily utilized for broadening the genetic base of cultivated lentil gene pool for improving the yield as well as wider adaptation.  相似文献   

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