山葡萄性别相关AFLP标记筛选及SCAR标记转化

通过AFLP分子标记方法,应用64对引物组合EcoR Ⅰ-NNN/Mse Ⅰ-NNN对山葡萄雌花、雄花和两性花植株基因池进行筛选,引物E-AGC/M-CTG在雄花和两性花植株基因池DNA中扩增出一条分子量为283 bp的特异性条带;经组池个体和其它品系验证,该特异性条带能在雄株和完全植株中稳定出现.将该特异性条带回收、克隆、测序,设计SCAR标记引物,对已知性别的85份山葡萄种质进行盲测,准确率达到97.6%,表明标记转化成功.由于该SCAR标记在绝大多数山葡萄栽培品种和有育种价值的种质资源中得到验证,可利用该SCAR标记对山葡萄杂交后代在苗期进行性别鉴定,提高育种效率.     

山葡萄性别相关AFLP分子标记的筛选及SCAR标记的转化

 山葡萄是一种重要的抗寒、抗病葡萄种质资源。通过AFLP分子标记方法,应用64对引物组合EcoR I-NNN/Mse I-NNN对山葡萄雌花、雄花和完全花植株基因池进行筛选,引物E-AGC/M-CTG在雄花和完全花植株基因池DNA中扩增出一条分子量为283 bp的特异性条带;经组池个体和其它品系验证,该特异性条带能在雄株和完全植株中稳定出现。将该特异性条带回收、克隆、测序,设计SCAR标记引物,对已知性别的85份山葡萄种质进行盲测,准确率达到97.6％,表明标记转化成功。由于该SCAR标记在绝大多数山葡萄栽培品种和有育种价值的种质资源中得到验证,可利用该SCAR标记对山葡萄杂交后代在苗期进行性别鉴定,提高育种效率。     

芦笋超雄株的DNA 分子标记辅助筛选

 以芦笋两性株自交S₁代雄株为试验材料,首先与优良雌株进行测交,然后利用与芦笋性别决定基因紧密连锁的显性DNA分子标记对苗期各单株进行性别鉴定,推断出超雄株,为芦笋全雄育种提供亲本材料。研究结果表明：显性标记Asp1-T7和Asp1-T7sp特异性好,扩增效果稳定,均能够鉴别芦笋雌、雄性别;一共测验了14株芦笋两性株自交S₁代的雄株,从中筛选出2株超雄株;对田间测交组合单株显花后性别表型调查的结果表明,DNA分子标记检测结果与田间实际相符,2株超雄株所对应的组合被确证为全雄组合。     

中国部分兰花品种RAPD分析

用随机引物扩增多态DNA（RAPD）分析技术，对中国兰花迸行品种鉴定和分类研究。使用10种20个碱基的引物，对19个中国兰花品种的基因组DNA进行扩增，其产生83条扩增带，其中80条大多态性带。每个兰花品种都有其特有的扩增带可与其它品种区别开来。这种特异的条带可作为一个品种的分子标记应用于品种鉴定和分类RAPD对基因组的分析结果与传统分类学的结果基本相符。各个种内的不同品种问亲缘关系接近，而不同种间差异较丸RAPD技术为中国兰花品种的鉴定和分类提供了新的途径     

番木瓜雄性性别的RAPD和SCAR标记

利用RAPD标记技术鉴定番木瓜雄性性状。在214条随机引物中,用引物Z18扩增得到1条雄性多态性片段,此片段存在于试验所用材料的所有雄株中而两性株和雌株没有,将其命名为Z18-1000。根据对该片段的序列分析结果重新设计了2对引物将RAPD标记成功转化成了SCAR标记,并命名为SD-1000。     

香榧性别鉴定的RAPD和SCAR标记

为了对香榧幼苗进行早期的性别鉴定,利用RAPD标记技术对香榧品种中的雌性和雄性群体进行了分析。在300条RAPD随机引物中,用引物S250扩增得到了1条大约600bp雄性特异性片段,将其命名为S250-600。进一步对该RAPD标记片段进行回收、克隆和测序,获得了593bp的该标记的核苷酸特定序列。根据对该片段的序列分析结果,设计了1对引物,成功地将RAPD标记转化为SCAR标记,命名为SW-593。将该序列递交至GenBank数据库,接收号为:EU670673。利用该SCAR标记对香榧的雌性和雄性个体进行性别鉴定,结果表明,该SCAR标记在雄性个体中均能稳定出现,说明此SCAR标记是一个与香榧雄性基因连锁的标记,可以用来对香榧品种进行大规模的性别鉴定。     

基于RAPD分析的李子新品种鉴定

以改进CTAB法对玉皇李和未鉴定新品种叶片基因组DNA的提取进行了试验。结果表明,该方法从这2个品种李树叶片中均能提出基因组总DNA,且DNA的纯度和完整性均较好,OD_(260)/O D_(280)的比值在1.8～2.0之间,无降解现象,RNA去除干净;此外,运用RAPD标记方法对这2个李品种基因组总DNA进行了PCR扩增,其RAPD分析条带清晰,用20条随机引物共扩增出114条带,平均每条引物扩增出5.7条条带,扩增片段长度为200～2000 bp,其中P_6、P_7、P_(10)、P_(14)和P_(18)5条随机引物可以单引物区分这2个品种,共获得20个条带,其中差异条带有6个,表明这6个位点存在差异。因RAPD每条扩增谱带对应基因组DNA分子上的一个位点,说明供试品种间DNA位点存在差异,表明未鉴定新品种在栽培过程中发生突变和分化,可能是1个优良种源。     

阿月浑子性别鉴定的形态学观测法研究

以阿月浑子雌、雄株的叶片、枝条为试材，进行叶形指数、成枝力及叶绿素吸光值的调查和统计，结果表明，无论是已知性别的成龄阿月浑子，还是未知性别的幼龄阿月浑子，其雌、雄株叶形指数间均表现显著差异；成枝力仅在成龄阿月浑子雌、雄株间差异非常显著；而叶绿素吸光值在成年及幼龄阿月浑子雌、雄株间差异均不显著。     

十八个麝香百合基因型遗传差异性的RAPD分析

对18个麝香百合基因型的遗传差异性进行了RAPD分析,筛选具有优良性状的新品种,并尝试使用RAPD标记技术的DNA指纹图谱作为DUS测试的分子生物学依据.结果表明:14条引物共扩增出142条带,其中多态性条带106条,多态百分比为74.65%,总体多态百分比不是很高,说明各基因型之间遗传差异较小.对RAPD扩增结果进行聚类分析,这18个基因型可以分成4类.一些引物扩增的结果显示,部分基因型具有特异性条带,说明RAPD技术在DUS测试中可以从分子生物学性状来鉴定新品种的特异性.     

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。     

13个阿月浑子品种的RAPD分析

为探明引进的阿月浑子品种间的亲缘关系,采用RAPD 标记技术对引入的12 个国外品种和‘新疆优株’进行了分析。用筛选的21 条10 bp 随机引物进行PCR 扩增可扩增出137 个位点,其中多态位点122 个,多态位点比率89.05%;品种间遗传距离在0. 2015 ~ 0.5163 之间,表明各品种间存在一定的遗传差异。UPGMA 聚类分析表明,13 个阿月浑子品种在遗传距离0.40 处可划分为3 个类群,第1 类包括 ‘Kerman’、‘Larnaka’、‘M-38’、‘M-P3’、‘Ashoury’、‘N’、‘Joley’、‘Aegina’、‘Avidon’、‘B’和‘Peter’11 个品种;第2 类为‘Xinjiang select tree’品种;第3 类为‘Mateur’品种。     

Development of sex-associated RAPD markers in wild Pistacia species

Summary

Marker assisted selection may greatly facilitate pistachio rootstock breeding as well as cultivar improvement, because of the long juvenile period of Pistacia species. Early diagnosis of seedling sex type would assist breeding and nurserymanagement in these dioecious species. We searched for RAPD markers linked to sex in P. atlantica, P. terebinthus and P. eurycarpa, the main wild species in Turkey that are used as rootstocks for P. vera. For this purpose, leaf samples were collected from male and female individual trees from each species and sex-pooled DNA samples were prepared by mixing the DNA of ten male and ten female individuals, to screen for sex associated RAPD bands. A total of 472 primers have been screened so far and two bands, amplified by primers BC156 and BC360, appeared to be sex assocaited in P. eurycarpa. The bands were tested in 30 male and 37 female individuals. Band BC156 (1300) was present in all, except one, female trees and was absent from all the male trees. Band BC360 (500) was amplified in 31 out of 37 females and was absent from all the males. In P. atlantica, one primer, OPAK09, amplified a femaleassociated band (850 bp), that was present in all 46 female individuals tested and absent in all the 38 male trees tested. It is likely that these markers are linked to sex-determining loci. The sex determination mechanism has not been characterized in Pistacia and segregating populations from controlled crosses are required to elucidate such mechanism and also to measure the genetic distance of our markers from the putative sex loci.     

我国阿月浑子生产现状及发展对策

介绍了阿月浑子（PistaciaveraL．）的原产地及分布，我国的栽培现状和存在的问题。提出了我国发展阿月浑子生产的对策：1．引进国外优良品种，丰富我国种质资源；2．注重当地优良品种的选育；3．扩大引种试点，扩大引种范围，尽快划定阿月浮子适生区域；4．学习国外阿月浑子生产的先进经验，向科研和生产的广度迈进。     

阿月浑子结果特性研究现状

介绍了国外阿月浑子大小年结果的研究进展,包括以下几个方面：（１） 花序芽分化及其脱落;（２） 树体对大小年结果的响应;（３） 激素与花序芽脱落;（４） 大小年结果的控制措施;（５） 大小年结果机理和存在的问题     

Analysis of genetic diversity among Indian short-day onion (Allium cepa L.) cultivars using RAPD markers.

Summary

Randomly amplified polymorphic DNA (RAPD) markers were used to estimate genetic diversity among 24 cultivars of short-day onions. Total genomic DNA was extracted and subjected to RAPD analysis using 90 arbitrary 10-mer primers. Of these, 15 primers were selected which yielded 137 bands, 91.24% of which were polymorphic. None of the primers produced a unique banding pattern for each cultivar. RAPD data were used to calculate a Squared-Euclidian Distance matrix which revealed a minimum genetic distance between cultivars ‘AFLR-722’ and ‘PBR-140’, and a maximum genetic distance between cultivars ‘PBR-139’ and ‘A.Kalyan’, and ‘MS-48’ and ‘A.Kalyan’. Based on the distance matrix, cluster analysis was done using a minimum variance algorithm.The dendrogram thus generated, based on Ward’s method, grouped the 24 onion cultivars into two major clusters. The first cluster consisted of cultivars from the northern region, and the second of cultivars from the southern region of India. The present study shows that there is high diversity among the onion cultivars selected and indicates the potential of RAPD markers for identification and maintenance of onion germplasm for crop improvement purposes.     

梨属种质资源的RAPD分析

 对梨属40 个品种及类型进行了RAPD 扩增, 根据相似系数进行UPGMA 法聚类,将起源于欧洲的西洋梨与起源于中国的3 个种明显区分开, 起源于中国的3 个种虽未完全各自聚类, 但各材料间都有不同的遗传距离, 证明RAPD 技术可作为梨种质分类鉴定的依据。     

土壤含水量对阿月浑子叶片净光合速率及叶绿素荧光参数的影响

 研究了土壤相对含水量对阿月浑子叶片叶绿素含量、净光合速率(Pn) 和荧光参数的影响。结果表明: 干旱胁迫降低了阿月浑子叶片Chl1a、Chl1b 含量和Pn , 增加了类胡萝卜素含量; 20 %供水处理降低了阿月浑子叶片的PS Ⅱ活性, 原初光能转化效率(Fv/ Fm) , 光化学猝灭系数(qP) , 非光化学猝灭系数(qN) 和PS Ⅱ电子传递量子产量(ΦPS Ⅱ) 。轻度干旱胁迫下, 气孔限制是Pn 降低的主要原因; 严重干旱胁迫下, 非气孔限制是Pn 降低的主要原因。干旱胁迫下阿月浑子叶片Pn 的下降是水分胁迫和强光胁迫双重作用的结果, PS Ⅱ电子传递量子产量(ΦPS Ⅱ) 降低是阿月浑子叶片Pn 下降的内在原因。     

RAPD技术在杨梅属植物分类研究中的应用

对杨梅属植物的２４个材料进行了ＲＡＰＤ型带及聚类分析。结果表明：ＲＡＤ技术能将起源于北美中心的蜡杨梅与起源于中国的３个种明显区分开，起源于中国的３个种各自聚类，各材料之间都有不同的遗传距离，证明ＲＡＰＤ技术可作为种级水平的分类鉴定依据。对ＲＡＰＤ技术在杨梅属植物分类研究中的问题进行了探讨。     

Effect of pistil age and pollen parent on pollen tube growth and fruit production of pistachio

Female pistachio (Pistacia vera) trees were hand pollinated with pollen from three different male genotypes 0,1,2,3 and 4 d after anthesis. Results were measured by mature fruit set and by fluorescence microscopy of pollen germination and tube growth in the pistil. Maximum pollen tube growth and fruit set of split nuts was achieved following pollination within two days of anthesis. Three day old pistils supported pollen tube growth, but fruit set was low. Four day old pistils supported little pollen tube growth or fruit set. Significant differences were also apparent between pollen parents, with low pollen tube growth and fruit set following interspecific pollination with P. atlantica pollen. All pollen tubes were observed to penetrate chalazogamously, and parthenocarpic production of blank fruits occurred in both unpollinated and pollinated treatments. The results indicate that for optimum fruit production of pistachio, pollen transfer must be achieved within 2 d of anthesis and pollination by P. atlantica should be avoided.     

Assessment of genetic relatedness among mango cultivars of India using RAPD markers

Summary

DNA-based RAPD (Random Amplification of Polymorphic DNA) markers have been used extensively to study genetic relationships in a number of fruit crops. A wide genetic diversity exists in the mango fruit in India. Present day commercial cultivars originated mainly from this subcontinent. In this study, 18 commercial mango cultivars, traditionally grown in western, southern, northern and eastern parts of India, were selected to assess genetic relatedness. Total genomic DNA was extracted and subjected to RAPD analysis using 30 arbitrary 10-mer primers. Of these, 27 primers amplified mango genomic DNA. None of these primers produced unique band pattern for each cultivar. RAPD data were used to calculate a squared Euclidean distance matrix, and based on this cluster analysis was done using a minimum variance algorithm. Cluster analysis clearly showed two groups—the first consisting of western, northern and eastern mango cultivars and the second group consisting of southern cultivars. From the analysis of results, it appears the majority of mango cultivars originated from a local mango genepool and were domesticated later.