SSR标记鉴定甜瓜品种‘红月亮’种子纯度

为明确甜瓜品种‘红月亮’F1代的种子纯度,通过提取甜瓜‘红月亮’子叶DNA,利用SSR(Simple Sequence Repeats)分子标记对甜瓜品种红月亮F1代种子DNA进行了PCR扩增、检测与分析。结果表明,在20个SSR引物组合对亲本及F1代筛选中,共有6对引物组合具有多态性,多态率30%。选择CMBR026用于‘红月亮’F1代种子纯度鉴定,使用该标记共检测118株,其中2株为自交株,种子纯度为98.3%;试验用的‘红月亮’种子田间鉴定纯度为99.5%,鉴定结果一致。SSR分子标记方法更能准确鉴别出‘红月亮’中的不纯株,且大大缩短纯度鉴定周期。     

粤椒1号辣椒种子纯度的RAPD检测研究

在分子标记鉴定蔬菜作物种子纯度的理论体系和技术体系指导下，对粤椒1号辣椒种子纯度进行了RAPD检测研究。结果表明，通过对200余个随机引物的筛选，发现特异引物P1、P2可应用于粤椒1号辣椒种子纯度的检测；对1份种子样品进行了RAPD检测，其结果与田问种子纯度检测结果完全一致。     

SSR标记鉴定甜瓜品种‘红月亮’种子纯度

为明确甜瓜品种‘红月亮’F1代的种子纯度,通过提取甜瓜‘红月亮’子叶DNA,利用SSR（SimpleSe—quenceRepeats）分子标记对甜瓜品种红月亮F。代种子DNA进行了PCR扩增、检测与分析。结果表明,在20个SSR引物组合对亲本及F。代筛选中,共有6对引物组合具有多态性,多态率30％。选择CMBR026用于‘红月亮’F1代种子纯度鉴定,使用该标记共检测118株,其中2株为自交株,种子纯度为98．3％;试验用的‘红月亮’种子田间鉴定纯度为99．5％,鉴定结果一致。SSR分子标记方法更能准确鉴别出‘红月亮’中的不纯株,且大大缩短纯度鉴定周期。     

SRAP标记在夏华2号甘蓝种子纯度检测中的应用

应用SRAP技术对夏华2号甘蓝种子纯度进行了分子检测研究。结果表明,从100对SRAP引物中筛选出1对引物(Me-5/Em-8)能清晰地扩增出父、母本的特异性条带,并且F1代带型呈双亲互补。利用这对SRAP引物对夏华2号种子进行纯度检测,检测纯度为97%,与田间生物学性状表现相符合,表明SRAP技术可用于夏华2号甘蓝种子纯度鉴定。     

甜瓜分子标记纯度鉴定研究

以2个甜瓜品种西域雪1号、西域雪2号及其亲本为试材,建立并优化了单粒种子DNA快速提取法和适合甜瓜的RAPD、SSR分子标记杂交种纯度鉴定体系。从100条随机引物中筛选出G17用于西域雪2号纯度鉴定,可有效区分母本和杂交种。从19对SSR引物中筛选出9对稳定扩增、多态性丰富的引物构建西域雪1号、西域雪2号的SSR指纹图谱。SSR分子标记用较少引物即可扩增出品种特异性条带,且多数F1代带型呈双亲互补,非常适合种子纯度鉴定。同时,使用高浓度、非变性的聚丙烯酰胺凝胶电泳、银染,条带更清晰,可区分只相差1个碱基的扩增片段,同RAPD相比,SSR分子标记更适合甜瓜杂交种纯度快速鉴定。     

结球甘蓝西园4号种子纯度的SSR鉴定

利用SSR分子标记技术对结球甘蓝西园4号及其亲本的DNA进行扩增,从90对SSR引物中筛选出1对具有双亲条带差异明显的引物O112-G04,构建了清晰的DNA扩增指纹图谱,并对西园4号种子进行了纯度鉴定,与田间形态学的鉴定结果高度一致,种子纯度均为96.7 %。结果表明,利用这对引物对结球甘蓝西园4号种子的纯度鉴定是可行的。     

结球甘蓝西园4号种子纯度的SSR鉴定

利用SSR分子标记技术对结球甘蓝西园4号及其亲本的DNA进行扩增,从90对SSR引物中筛选出1对具有双亲条带差异明显的引物O112-G04,构建了清晰的DNA扩增指纹图谱,并对西园4号种子进行了纯度鉴定,与田间形态学的鉴定结果高度一致,种子纯度均为96.7%.结果表明,利用这对引物对结球甘蓝西园4号种子的纯度鉴定是可行的.     

利用SSR技术鉴定西瓜甜瓜种子纯度

利用SSR分子标记技术对西瓜‘W1806’与甜瓜‘M1805’各3个批次及其亲本间的多态性进行引物筛选和种子的纯度鉴定。结果表明,在28对西瓜的SSR引物中,有5对引物在西瓜‘W1806’的F1代与亲本之间有很好的多态性。其中BVWS00839引物特异性好,条带清晰,父母本条带间隔明显,即作为3个批次的西瓜纯度鉴定的引物,其鉴定纯度分别为99.47%、98.96%和97.92%;在18对甜瓜的SSR引物中,只有1对CMBR052引物在F1代扩出的条带为典型的双亲互补型条带,故用该引物对甜瓜进行纯度鉴定,其纯度分别为97.90%、96.80%和97.40%。与田间鉴定结果的吻合率都在98%以上。这2个材料的吻合率说明SSR分子标记技术对西瓜和甜瓜纯度鉴定结果都是可靠的。     

14个辣椒杂交种纯度鉴定的SSR引物筛选

利用73对多态性较好的辣椒SSR引物,对湘研种业的14个辣椒品种进行差异性筛选,结果表明,每个品种都有1对以上SSR引物可将其父本、母本及F1代区分开;同时对每个品种的多个批次进行验证,检测结果与田间检测结果基本相符。因此,可利用SSR分子标记技术进行辣椒种子室内纯度鉴定,且具有简单、快速、准确、重复性好等优点,SSR分子标记在辣椒种子室内纯度快速检测方面具有很大的应用前景。     

利用SSR分子标记鉴定“中丝2号”丝瓜杂交种纯度

为了建立一套丝瓜种子纯度的快速鉴定方法，采用SSR分子标记对丝瓜杂交一代品种“中丝2号”的纯度进行鉴定，利用24对SSR引物对“中丝2号”丝瓜及其亲本的DNA进行扩增。结果表明：从24对引物中筛选出1对特异性引物ZS-SSR24，能够分别在“中丝2号”母本和父本中扩增出特异的2条多态性条带，大小分别为220 bp和250 bp,F1继承双亲的特异性条带，利用此标记能够鉴定出混在F1中的父本或母本。经田间鉴定，供试丝瓜种子纯度为91.3%，这与分子标记的结果完全吻合，故该标记能够用于“中丝2号”丝瓜品种的纯度鉴定。     

Assessment of genetic purity of tomato (Lycopersicon esculentum L.) hybrid using molecular markers

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.     

Genetic diversity of ash gourd [Benincasa hispida (Thunb.) Cogn.] inbred lines based on RAPD and ISSR markers and their hybrid performance

Ten inbred lines of ash gourd [Benincasa hispida (Thunb.) Cogn.] were crossed to produce 45 F₁ hybrids (without reciprocal) which were evaluated along with the parents for 20 growth- and yield-related traits, in a replicated field trial. High level of heterosis was observed among the hybrids for most of the traits examined, including yield. These inbred lines were analysed by using 42 RAPD primers those produced 282 DNA marker bands. A total of 130 RAPD markers were obtained with a mean of 3.1 per primer, which in combination discriminated all the inbreds from each other. Pair-wise genetic distance measurements ranged from 0.07 to 0.31, suggesting a wide genetic diversity for these inbreds. These inberds were also analysed with five ISSR primers of which four were informative. Twenty-six ISSR marker bands were generated of which 11 were polymorphic with an average of 2.80 per primers. The percentage of polymorphic bands produced were higher in ISSR markers (>80%) than generated through RAPD markers (46%). Although the results indicated significant positive correlations of genetic distance with hybrid performance and heterosis, the RAPD based genetic distance measures and use of limited ISSR markers in this present study could not effectively predict hybrid performance in this crop. The genetic variation among ash gourd inbred lines examined, herein, defined a marker array (combined ISSR and RAPD) for the development of a standard reference for further genetic analyses, and the selection of potential parents for predicting hybrid performance and heterosis.     

利用RAPD技术快速鉴定辣椒杂交种纯度的研究

以新椒系列辣椒杂交种父母本和一代杂种为材料,采用液氮-SDS法提取辣椒基因组DNA,研究PCR反应的主要影响因子,建立适合辣椒杂交种纯度RAPD分析的PCR反应体系,从120个随机引物中筛选出1个能鉴定新椒10号杂种纯度的引物S1220,从7个随机引物中筛选出1个能鉴定新椒3号杂种纯度的引物S1213。     

Comparative analysis of genetic diversity in Indian bitter gourd (Momordica charantia L.) using RAPD and ISSR markers for developing crop improvement strategies

A genetic analysis of 38 diverse Indian bitter gourd (Momordicacharantia var. charantia, and var. muricata) accessions was performed using 29 RAPD and 15 ISSR markers. RAPD primers yielded 208 amplicons of which 76 (36.5%) were polymorphic providing an average of 2.6 amplicons per primer. RAPD amplicons per primer ranged from 3 (OPE-19, OPW-09) to 15 (OPW-05), and varied in size from 200 bp to 3000 bp. Fifteen ISSR primers provided a total of 125 bands of which 94 (74.7%) were polymorphic. Polymorphic ISSR markers ranged from 0 (UBC-841) to 12 (UBC-890) providing a mean of 6.3 amplicons per primer that ranged in size from 150 bp to 2700 bp. Nevertheless, the concordance among bitter gourd accession groupings after cluster analysis was relatively high (r = 0.77), indicating that RAPD- and ISSR-based diversity assessments in this germplasm array were generally consistent. The M.charantia var. charantia (domesticated) and var. muricata (wild, free-living) accessions examined were genetically distinct, and these differences provided for the development of strategies for genetic analyses and crop improvement in this species.     

Genetic relatedness (diversity) and cultivar identification by randomly amplified polymorphic DNA (RAPD) markers in teasle gourd (Momordica dioica Roxb.)

Genetic relationship and variation of 29 accessions of teasle gourd (Momordica dioica Roxb.) and 1 accession of Momordica cochinchinensis Spreng. (wild relatives of teasle gourd) were examined by RAPD analysis using 44 dodecamer oligonucleotide primers. A total of 496 fragments were produced by 44 primers of which 95% bands were polymorphic. Using presence or absence of specific RAPD markers or combination of primers, 23 out of 30 accessions were identified. The genetic relatedness or genetic distance based on Nei and Li's genetic similarity varied from 0.86 to 0.65 with an average of 0.74 among 29 M. dioica accessions (when M. cochinchinensis excluded). In the phenetic dendrogram developed from cluster analysis using UPGMA method, M. cochinchinensis was out grouped as single accession, while others showing relatively weak grouping formed four groups. Clustering pattern did not demonstrate any relationship between geographical origin and genetic diversity. A DNA extraction method has been standardized. This is the first report of using RAPD techniques in teasle gourd. It was concluded that RAPD analysis is a useful tool for genotypic identification and estimation of genetic similarity in teasle gourd.     

Conversion of chromosome-specific RAPDs into SCAR-based anchor markers for onion linkage maps and its application to genetic analyses in other Allium species

Integration of previously developed Allium cepa linkage maps requires the availability of anchor markers for each of the eight chromosomes of shallot (A. cepa L. common group Aggregatum). To this end, eight RAPD markers originating from our previous research were converted into SCAR markers via cloning and sequencing of RAPD amplicons and designing of 24-mer oligonucleotide primers. Of the eight pairs of SCAR primers, seven resulted in the amplification of single bands of the original RAPDs, and the remaining primer set amplified an additional band. The results of Southern hybridization using RAPD amplicons from genomic DNA of Japanese bunching onion (Allium fistulosum L.)—shallot monosomic addition lines indicated that five SCAR markers were single shallot chromosome-specific markers and were not detected in genomic DNA of A. fistulosum. The eight SCAR primer pairs were applied to other Allium species and exhibited three types of amplification profiles, namely RAPD amplicons observed only in shallot, in shallot and Allium vavilovii, and in several Allium species. A mapping study using 65 F₂ plants generated by the selfing of one interspecific cross A. cepa × Allium roylei individual integrated the SCAR marker SAOE17₅₀₀ into chromosome 5 as expected. The results of the present study show that the eight SCAR primer sets specific to shallot can facilitate the mapping in A. cepa and can also serve as anchor points between maps of different Allium species.     

盐溶液引发对冬瓜种子活力及其生理变化的影响

用10 mmol·L^-1的CaCl₂、NaCl、KNO₃、KH₂PO₄、CaCl₂+NaCl（1∶1）、KNO₃+KH₂PO₄（1∶1）溶液分别对黑皮冬瓜种子进行引发处理,探究盐溶液引发对冬瓜种子活力及生理变化的影响。结果表明：10 mmol·L^-1 KNO₃溶液处理能显著提高冬瓜种子的活力,促进种子萌发,种子发芽势、发芽率、简化活力指数、SOD和CAT活性分别提高4.0 %、14.7 %、60.0 %、53.2 %和78.7 %,电导率降低51.2 %。     

辣椒RAPD反应体系建立及杂交种纯度鉴定研究

以豫椒系列品种父母本和杂交种为材料，采用液氮-SDS法提取辣椒基因组DNA，使用Bio-RAD-Mycy-cler型PCR仪和Bio-RAD1000凝胶扫描仪，研究PCR反应的主要影响因子，建立适合辣椒RAPD分析的PCR反应体系，从40个10bp的随机引物中筛选出1个能鉴定豫椒968杂种纯度的引物S149。     

冬瓜和节瓜种质资源遗传多样性的RAPD分析

采用RAPD分子标记技术对41份冬瓜和节瓜种质资源进行遗传多样性分析。从455条随机引物中筛选出23条能产生稳定多态性扩增谱带的引物用于RAPD分析,共产生143条扩增谱带,其中多态性谱带81条,多态性检测率为56.6%,表明冬瓜和节瓜种质资源在分子水平上具有丰富的遗传多样性。用NTSYSpc软件处理RAPD数据,41份材料间的遗传相似系数在0.60～0.99之间。聚类分析结果可将供试材料分为6大类群9个亚类,主坐标分析可将供试材料分为6大类群10个亚类。两种分类方法的结果基本一致。