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1.
Anthracnose caused by the fungus Colletotrichum lindemuthianum is the most destructive disease of cowpea. Field-type cowpeas show various levels of resistance, whereas pole-type vegetable cowpeas are highly susceptible. Transfer of resistance available in field types to vegetable types is a major breeding objective in cowpea. This paper details the development of an F2 mapping population by crossing field-type cowpea variety Kanakamony (Vigna unguiculata ssp. cylindrica) with pole-type vegetable cowpea variety Sharika (Vigna unguiculata ssp. sesquipedalis), screening this population with artificial inoculation and Bulked Segregant Analysis (BSA) with random marker systems Random Amplified Polymorphic DNA (RAPD) and Inter-Simple Sequence Repeats (ISSR); the objective is to identify the markers linked with major resistance-contributing genes. RAPD primer OPA02 has yielded marker at 850 bp in susceptible genome, whereas ISSR primers UBC810 and UBC811 have yielded markers at 1.4 kb and 1.5 kb respectively in resistant genomes. These markers were reproducible and their linkage with resistance and suitability in marker assisted selection (MAS) were confirmed through co-segregation analysis in F3 population. UBC811 marker was eluted, cloned on pGEM-T, and sequenced. The sequence had shown that this marker is anchored on LRR receptor-like serine/t\hreonine protein kinase gene which could be involved in the resistance mechanism.  相似文献   

2.
用于黄瓜白粉病抗病性鉴定的SCAR标记   总被引:2,自引:1,他引:1  
将黄瓜白粉病抗性相关基因的一个共显性AFLP标记成功地转换为简单、实用的SCAR标记。根据AFLP标记片段序列信息 设计了两对特异性引物SCPM166和SCPM66。PCR扩增结果显示,SCPM166在抗病亲本、抗病单株和杂合类型单株中均扩增出了大 小为166 bp的特异片段,感病亲本与感病单株均无此特异片段,表明该SCAR标记可以鉴定纯合感病个体;SCPM66在感病亲本 、感病单株和杂合类型单株中均扩增出了大小为66 bp的特异片段,抗病亲本与抗病单株均无此特异片段,表明该SCAR标记可 以鉴定纯合抗病个体。若将两个显性SCAR标记结合起来,相当于一个共显性的SCAR标记。  相似文献   

3.
The root-knot nematodes (Meloidogyne spp.) are important nematode pests and cause serious diseases in pepper in the world. No molecular markers linked to the nematodes resistance N gene have been reported. In this paper, ‘Carolina Wonder’ (Capsicum annuum L.), a sweet pepper line resistant to root-knot nematode with N gene, ‘20080-5-29’ (C. annuum L.), an inbred line susceptible to root-knot nematode with good horticultural characteristics, and their F2 progeny with 320 individuals were used as materials. Evaluation of resistance and susceptibility of parental lines, F1 and F2 progeny inoculated with root-knot nematodes (Meloidogyne incognita) were carried out. ‘Bulked segregant analysis’ method was used to search for polymorphic markers from 512 pairs of AFLP primers. Based on the assessment of resistance and susceptibility and polymorphism of the AFLP marker in F2 population, the genetic linkage distance between the AFLP marker and the N gene was estimated. One AFLP marker E39/M41-339 was obtained and transferred to a SCAR marker amplifying a 315 bp DNA fragment linked to the N resistant allele and a 331 bp fragment linked to the N+ susceptible allele. The distance between the molecular marker and the nematodes resistance N gene is 6.3 cM. This research delivered a valuable tool for the marker assisted selection of nematodes resistance in pepper.  相似文献   

4.
抗病转录因子Pti4是从番茄中分离的一种能够调控番茄抗病基因Pto转录的蛋白质。通过根癌农杆菌介导法将抗病转录因子基因Pti4导入花椰菜无菌苗下胚轴,同时对转基因植株进行了PCR、PCR-Southern和Southern blot分子检测以及细菌性黑腐病和软腐病病菌接种试验。结果表明,在建立再生系统过程中外植体的最佳苗龄为5 ~ 7 d,培养基中激素组合为NAA 0.2 mg • L-1、6-BA 2.0 mg • L-1 时不定芽的诱导和分化效果最好。获得的35株阳性植株经分子检测证明抗病转录因子基因Pti4已被整合到花椰菜的基因组中。对其中26株接种病菌发现Pti4转化植株对细菌病害具有一定的抗性。  相似文献   

5.
番茄抗晚疫病基因Ph-3的RAPD标记   总被引:4,自引:1,他引:3  
 以番茄抗晚疫病材料CLN2037和感病材料T2-03杂交的F2分离群体的147个单株为试材, 通过抗病性鉴定, 选择高抗单株和高感单株分别建抗、感病池。采用BSA法筛选了230条RAPD引物, 其中引物CCPB272-03 (序列为GGTCGATCTG) 在抗、感病池扩增出多态性片段CCPB272-03740 , 通过F2单株验证后证明CCPB272-03740与抗晚疫病基因Ph-3紧密连锁, 遗传距离为518 cM。  相似文献   

6.
Cardamom lines, NKE 9 and NKE 12, resistant to Cardamom Mosaic Virus (CdMV) was crossed to two susceptible genotypes, viz., CCS 1 and RR 1 to determine the nature of inheritance of resistance. It was revealed from the results that the CdMV resistance in NKE 9 and NKE 12 is genetically governed. The F1 hybrids between resistant and susceptible genotypes were resistant. The segregation pattern for disease reaction in F2 and BC1 generations of the two crosses suggested that CdMV resistance in NKE 9 and NKE 12 could be controlled by two dominant complementary genes. Over all it could be hypothesized that the resistance to CdMV is quantitative, with possibly two major factors, and dependant on gene dosage with completely dominant gene action. This is the first report of CdMV inheritance in cardamom.  相似文献   

7.
茄子青枯病抗性基因的遗传分析及其AFLP标记   总被引:8,自引:0,他引:8  
 以茄子高感青枯病品种‘三月茄’与高抗品种‘S69’的F1、F2为材料, 对青枯病抗性遗传进行了分析, 同时开展了与抗性基因连锁的AFLP标记鉴定。结果表明: ‘S69’对青枯病的抗病性属于不完全隐性遗传, 感病性属于不完全显性; 抗性由1~2对基因控制。对86个F2单株AFLP分析, 鉴定出1个与‘S69’抗性基因连锁的AFLP标记39A980 (该引物组合为E-ACC /M-CTG) , 该标记与抗性基因间的交换值为4.65% , 遗传距离为4.9 cM。  相似文献   

8.
白菜抗霜霉病基因的RAPD标记   总被引:8,自引:0,他引:8  
研究了与抗霜霉病基因紧密连锁的RAPD分子标记, 利用白菜抗病品种014和感病品种010杂交的F2群体144个单株为材料, 通过人工接种鉴定, 采用高抗单株和高感单株分别构建抗、感病池, 利用BSA法筛选了560条RAPD引物, 其中引物AY12在抗感病池中扩增出多态性片段AY121238 , 通过F2单株验证后证明AY121238与抗霜霉病基因紧密连锁, 其遗传距离为6.7 cM。  相似文献   

9.
The genetics of resistance to tomato yellow leaf curl virus (TYLCV) were studied in TYLCV-resistant lines developed by crossing wild species of Lycopersicon pimpinellifolium, L. hirsutum and L. peruvianum resistant to TYLCV with susceptible L. esculentum cv. Special Back. Crosses between TYLCV-resistant lines derived from the same wild species produced progenies similar to their parents in their level of resistance. However, progenies from interspecific crosses showed greater resistance than either parent suggesting that the genes for TYLCV resistance contributed by different wild species are probably not the same (non-allelic). The gene action for TYLCV resistance also varied with the source of resistance. Analysis of F1, F2 and BC populations for lines derived from L. pimpinellifolium showed that resistance to TYLCV in these lines is a quantitative trait with some dominance.  相似文献   

10.
Three DNA molecular marker systems, RAPD, ISSR and SSR, were used to test seed genetic purity of two commercial hybrid tomato (Lycopersicon esculentum L.) cultivars ‘Hezuo 903’ and ‘Sufen No. 8’. Genomic DNA from the two F1 hybrid cultivars and their corresponding parental lines was screened with 218 RAPD decamer primers, 54 ISSR primers and 49 SSR primers. Among the 321 primers, 4 primers for ‘Hezuo 903’ and 3 for ‘Sufen No. 8’, which could produce both female and male parent-specific markers, were selected for testing the genetic purity. A total of 210 hybrid individuals of each cultivar were analyzed using the identified primers. The combined results of the marker analysis showed that eight of the 210 F1 plants in ‘Hezuo 903’ and 13 of 210 in ‘Sufen No. 8’ were false hybrids, and the overall genetic purity of the two F1 hybrid seed lots was 96.2 and 93.8%, respectively. This study showed that RAPD and SSR markers could provide a practical and efficient tool in quality control of the tomato commercial hybrid seeds.  相似文献   

11.
Watermelon [Citrullus lanatus (Thunb.) Matsum. &; Nakai var. lanatus] is a cross-pollinating diploid species and is one of the most important dicotyledonous plants belonging to the family Cucurbitaceae due to its high nutritional value, rapid growth, sweet and juicy taste, and refreshing nature as a summer fruit. However, the insertion–deletion (InDel) markers available for watermelon cannot satisfy the demand to identify and map resistance genes in this major crop. In this study, we have characterised and developed numerous polymorphic InDel markers using Illumina re-sequencing of a male parental line that is resistant to Fusarium wilt. In total, 25,701 potential InDel markers were identified, with a mean size of 18 bp. From these, 7,387 potential InDel markers >?10 bp were selected and tested as candidate InDel markers to locate resistance genes for map-based cloning and polymorphism analysis. We designed and synthesised 69 primer pairs using Primer 3 software. Overall, 54 primer pairs detected polymorphism between the Fusarium-susceptible female parent ‘JSw01-1’ and the Fusarium-resistant male parent ‘JRw08-3’. Among the 69 InDel markers, 54 (78.3%) exhibited polymorphism at 179 PCR loci. Polymorphic information content (PIC) values ranged from 0.04 to 0.91, and the number of alleles per primer ranged from 1 to 11. The 54 InDel markers that were mined and exploited via Illumina re-sequencing of watermelon could accelerate the development of marker-assisted breeding and the progression of genetic research to understand resistance to Fusarium wilt in watermelon.  相似文献   

12.
花椰菜—黑芥体细胞杂交获得抗黑腐病异附加系新材料   总被引:1,自引:0,他引:1  
王桂香  严红  曾兴莹  盛小光  唐宇  韩硕  宗梅  陆坤  刘凡 《园艺学报》2011,38(10):1901-1910
 以花椰菜—黑芥体细胞杂种的自交及回交后代为材料,建立了结合抗病性人工接种鉴定,流式细胞仪DNA含量分析,B基因组特异分子标记和染色体原位杂交的大规模群体分析技术,在50份高代自交及回交材料中筛选获得了20份形态学特征偏向花椰菜,30份呈花椰菜—黑芥中间类型的材料。细胞DNA含量检测(FLC)及染色体计数结果表明,偏花椰菜的类型,DNA含量基本在2.24 pg · cell-1以下,染色体数在28条以下。经连续3年黑腐病病菌人工接种鉴定,获得高抗株系12个,抗性株系17个,占全部受试材料的14%。对其中3个(PFCN29 BC3-3-5、PFCN16-1 S2BC3-107和PFCN14-1 S1BC4-123)形态偏花椰菜且高抗黑腐病的植株进行基因组原位杂交,结果显示其分别为花椰菜染色体组附加了2、7和8条黑芥染色体的异附加系材料。这些植株可以用于后续的研究和作为育种桥梁材料,进一步获得黑腐病抗性的单体附加系或渐渗系。本研究表明通过体细胞杂交转育野生优异抗性资源,创新甘蓝类蔬菜育种种质的可行性。  相似文献   

13.
Of 200 genotypes of cauliflower screened for their reaction to blackrot disease (Xanthomonas campestris (Pam.) Dowson) ‘MGS-2⊕-3⊕’, ‘Pua Kea’ and ‘SN 445’, derived from Indian cauliflower cultivars, possessed a high degree of resistance. Neither ‘Snowball’ nor its relatives ‘Erfurt’ and ‘Alpha’ showed any resistance. For the plant breeder, soaking the seed in a bacterial suspension is the most useful screening-technique; less useful were the spraying of whole plants with a suspension and the severing of a leaf-tip beneath a suspension. There were positive and highly significant correlations between seed soaking and seedling reaction, seed soaking and adult plant reaction, and seedling reaction and adult plant reaction.  相似文献   

14.
Gynoecious is an important economic trait of cucumber for determinant of earliness and yield, yet genetic mechanism is not well understood for this trait. The experiment was conducted using F2 mapping population by crossing of PPC-2, a gynoecious and parthenocarpic line with Pusa Uday (monoecious and non-parthenocarpic cultivar). Out of 179 SSR markers screened, 39 markers differentiated the gynoecious and monoecious parents. However, only 17 markers were segregating with F2 mapping population, those were used for genotyping and linkage map analysis and these markers were placed along with F locus on chromosome 6 covering a total distance of 100.4cM. The SSR markers, SSR13251 and UW020605 were found to be closely linked to gynoecious (F) locus at 1.0 and 4.5 cM, respectively. The segregation of F2 population of PPC-2 × Pusa Uday and GPC-1 × Punjab Naveen and test crosses for sex type herein suggested that single dominant gene controlled the gynoecious sex expression in cucumber particularly in gynoecious genotypes PPC-2 and GPC-1. Therefore, the monogenic dominant nature of gynoecious sex identified in the present experiment and SSR markers closely linked to the F locus will be useful in marker-assisted backcross breeding for transfering gynoecious trait into horticulturally desirable varieties.  相似文献   

15.
 为了开发番茄对番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)抗病性鉴定的新技术,设计了在番茄试管苗中接种TYLCV的试验。将对TYLCV感病的品种‘Moneymaker’、‘中蔬6号’、‘丽春’和抗病品系‘AVRDC CLN 2123 A’在试管中进行组织培养,21 ~ 25 d后切取长2.0 ~ 2.5 cm的嫩枝,将嫩枝基部在含TYLCV-[CN:SH2]侵染性克隆质粒的农杆菌EHA 105菌株的悬浮液(OD600 0.5)中浸润1 min接种。14 d后,感病品种试管苗开始出现TYLCV症状并且愈来愈重,28 d后TYLCV症状十分典型,56d后症状极其严重:叶片窄小、黄化、极度卷曲,植株严重矮化,停止生长或死亡;而同样接种的抗病品系试管苗却无一发病。接种的番茄试管苗经PCR检测显示,从感病品种试管苗中均能扩增到356 bp的特异片段,而在抗病品系中无此条带。这些结果显示了在试管里鉴定番茄品种或材料对TYLCV抗性的可能性。  相似文献   

16.
Cashew is an important edible nut crop of tropics. Bulk segregant analysis (BSA) was carried out on DNA bulks constituted from F2 population and germplasm in order to link or associate molecular markers with economic characters. In all 458 RAPD, 31 ISSR and 21 pairs of SSR primers were used and identified polymorphic markers between parents. Though screening F2 bulks with these markers identified markers polymorphic between the bulks but none could be validated with the individuals of their bulks. Hence screening with germplasm bulks was carried out and could identify four RAPD markers polymorphic between the bulks for nut weight and plant stature and also between the individuals of their bulks. Of the four, three markers were associated with nut weight amplifying at 775, 475, 275, bp region in primers OPN 14, UBC 184 and UBC 185 respectively. Out of these three, two markers were specific to low nut weight and one marker was specific to high nut weight and their bands were present in greater frequency (50–77.8% and 75%) of individuals constituting the respective bulks. Similarly, the another marker UBC 185275 was detected which was specific to low plant stature and was present in 66.7% and 10% individuals constituting short and tall bulks respectively. Markers identified with bulks and with the individuals of bulks were validated further with more individuals of F2 and germplasm.  相似文献   

17.
Experiments were conducted to study genetics of fertility restoration and to examine distribution of RAPD markers (OPW19800 and OPP131400) linked with fertility restoration gene (Rf) in pepper (Capsicum annuum L.) inbreds. Forty-two hot and five sweet pepper inbreds were crossed on a cytoplasmic male sterile (cms) line CCA-4261 and F1s were evaluated for fertility restoration under open field conditions. DNA of 5 plants of CCA-4261 and individual plants of 47 inbreds was isolated and PCR reaction was performed using OPW19 and OPP13 primers. The results revealed that most of the hot pepper lines posses Rf gene. The Rf gene associated two markers, viz., OPW19800 and OPP131400 were not frequently distributed in the restorer inbred lines because presence of marker bands often does not coincide with the presence of Rf gene identified in many restorer inbreds. The case specific applications of both the RAPD markers have been described.  相似文献   

18.
Summary

Mume (Prunus mume Sieb. et Zucc.) and apricot (P. armeniaca L.) are similar in fruit and tree morphology, and exhibit high cross- and graft-compatibility with each other. It is therefore difficult to differentiate mume and apricot cultivars on the basis of morphological and phenotypical characteristics. Molecular markers were developed to differentiate nine mume from ten apricot cultivars. Four dominant, random amplified polymorphic DNA (RAPD) markers that can discriminate between mume and apricot cultivars (designated OPA15628, OPO10550, OPO20259, and OPU03415) were identified from 21 decamer primers. Two RAPD markers (OPO10550 and OPU03415) were developed into dominant sequence-characterised amplified region (SCAR) markers (SCO10 and SCU03). These SCAR markers could differentiate between all mume and apricot cultivars.  相似文献   

19.
The objectives of this study were to investigate genetic variation and relationships between Indonesia-, Australian- and European-based cultivars and to evaluate variation within Indonesia cultivars as all cultivars are open-pollinated. Eight cauliflower cultivars collected from three production regions in Indonesia and four F1 hybrids cultivars grown in Australia were evaluated using RAPD and ISSR markers. DNA polymorphisms generated from 10 polymorphic RAPD primers were used to construct a dendogram using the unweighted pair-group method with arithmetic averages (UPGMA) and to generate a fingerprinting key. ISSR marker analysis using 14 primers were attempted but DNA polymorphisms could not be clearly identified. The RAPD technique indicated that variation occurred both within and between Indonesian cultivars. Comparison between Indonesian-, Australian- and European-based cultivars showed that Indonesian cultivars have unique genotypes and would be good sources of genes for future crop improvement.  相似文献   

20.
茄子抗青枯病基因的RAPD标记研究   总被引:8,自引:1,他引:8  
 为探明茄子抗青枯病的遗传机制, 以高感青枯病品种北京六叶茄064, 高抗青枯病半栽培种马来西亚S3及其F2 代为材料, 用RAPD技术对供试材料的抗青枯病基因进行了研究。结果表明: 通过300个随机引物的PCR扩增分析, 发现15.6%的引物在双亲间表现出多态性, 找到了一个与茄子抗青枯病亲本S3中的抗病基因紧密连锁的分子标记S264780 (该引物的碱基序列为CAGAGCGGA) , 该标记与S3的抗病基因的交换值为4.32% , 遗传距离为4.33 cM。  相似文献   

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