西瓜DUS测试标准品种SSR指纹图谱构建及应用

本研究采用代表最大限度西瓜遗传多样性的SSR核心引物组合,分析了西瓜DUS测试指南中的24份标准品种遗传多样性与核酸指纹。以基于重测序获得的SNP标记构建的17份西瓜材料的系统发育树为参照,对24份标准品种进行了遗传多样性分析,在遗传相似系数0.80处将24份标准品种分为3大类群,分析表明：核酸指纹分类比传统形态学分类更为准确。采用二维（QR）编码构建了西瓜24份标准品种的SSR指纹图谱,并利用本技术以保护品种“京欣2号”与对照品种“京欣1号”为例,进行了DUS分子鉴定测试,共扩增32个SSR位点,“京欣1号”和“京欣2号”之间存在4个位点的差异,品种间遗传相似系数为0.89,比形态学鉴定的差异位点更多且更准。本研究建立的西瓜DUS标准品种SSR指纹图谱与分子检测技术,可以应用到西瓜品种DUS分子检测实践,同时也为西瓜品种纯度与真实性鉴定及遗传背景分析提供了技术方案。     

薏苡新品种特异性、一致性和稳定性测试指南研制

植物新品种满足特异性、一致性和稳定性(合称DUS)是授予品种权的前提条件之一。DUS测试指南是开展DUS测试的技术依据。本文详细介绍了薏苡DUS测试指南的研制过程,包括适用范围、繁殖材料的要求、测试性状的选择、性状表达状态和相应代码的确定、标准品种的筛选、DUS判定标准和技术问卷的设计等。薏苡指南的研制对促进中国薏苡新品种保护,鼓励品种创新及加强品种管理具有重要意义。     

武夷山及周边地区黄精植物ISSR分子标记鉴定及HPLC指纹图谱研究

采用ISSR分子标记鉴定方法及HPLC色谱方法对武夷山及周边地区5份黄精植物样本进行DNA分子标记鉴定及指纹图谱分析,并通过聚类分析软件分别探讨其遗传相关聚类图谱。ISSR分子标记法能较好地对5份黄精植物样本进行区分,HPLC指纹图谱分析同样表明5份黄精样本化学成分存在一定的差别。DNA分子标记及HPLC指纹图谱分析的遗传相关聚类分析结果相似,表明武夷山及周边地区的野生黄精种质资源存在明显的差别。     

无髯鸢尾新品种DUS测试性状筛选

植物新品种特异性、一致性和稳定性(DUS)测试是新品种保护的基础和依据,测试性状的筛选是DUS测试的核心。为制定无髯鸢尾DUS测试指南,初步确定测试性状43个,结合性状间R型聚类分析性状间相关性,去除相关性大、贡献较小的雌蕊长、叶长性状,最终确定操作性强、区别明显的测试性状41个;利用品种间Q型聚类分析确定测试指南的分组性状4个;对性状进行分级及分布分析,保证无髯鸢尾测试指南的可行性,为无髯鸢尾品种测试与授权提供技术支持。     

小麦DUS测试已知品种DNA指纹数据库构建及其应用

为解决DUS测试中特异性测试近似品种筛选难的问题,本研究基于2256份DUS测试小麦已知品种构建DNA指纹数据库。首先利用81对SSR引物对96份小麦品种进行遗传相似度分析。结果表明,81对SSR引物扩增出84个位点,共检测到731个等位变异,每个位点的等位变异数在3～18之间,多态性信息指数(PIC)变化范围为0.29～0.89。选取42个SSR标记构建了2256份小麦品种DNA指纹数据库,共获得148493个数据,除引物barc14、xgwm341之外,其他引物数据缺失率在5%以内。对176份小麦品种进行DNA指纹数据采集和特异性测试,结果表明不具有特异性的申请品种与其最近似品种的遗传相似度都在90%以上;本研究基于42对SSR荧光标记引物构建了我国小麦DUS测试已知品种的DNA指纹数据库,确定近似品种筛选的遗传相似度阈值为80%,建议将与申请品种遗传相似度高于80%的已知品种作为近似品种进行田间种植和特异性评价。     

水稻新品种测试的标准品种DNA指纹图谱多样性分析

以植物新品种特异性、一致性和稳定性(DUS)测试指南中选定的49个水稻标准品种为材料,采用农业行业标准(NY/1433-2077)中推荐的24对水稻SSR引物进行DNA指纹图谱构建和遗传多样性分析。结果表明,24对SSR引物在19个籼稻和30个粳稻品种中分别检测到141和156个等位变异,平均每对引物可以检测到5.88(籼稻)和6.50(粳稻)个等位变异;籼、粳稻类群中24对SSR引物的平均Shannon's多样性指数分别为1.5141(1.0460~1.9959)和1.4389(0.4677~2.4503);经聚类分析后,籼、粳稻群体内品种间的相似系数分别介于0.45~0.81和0.36~0.76,取相似系数0.72为阈值,可将19个籼稻和30个粳稻品种分别分为16类和28类。由此可见,本研究的49个品种的遗传多样性丰富。结合形态性状和基因型聚类分析结果,可将现有的49个水稻DUS测试标准品种减少到46个。     

应用SRAP标记绘制88份南瓜属种质资源DNA指纹图谱

为了给南瓜属种质资源鉴定和分类提供分子生物学依据,本研究采用SRAP分子标记技术与DNAMAN指纹图谱绘制软件对88份南瓜属种质资源(包含美洲南瓜、中国南瓜、印度南瓜)进行分子指纹图谱绘制。结果表明:35对SRAP多态性引物共扩增出499条清晰条带,其中多态性条带438条,多态性条带比率高达87.8%。根据扩增出的条带成功绘制出88份南瓜属种质资源的DNA指纹图谱,每一份种质都具有其独特的分子身份证,使得每份种质均可被区别开来。其中,多态性最好的引物是E5EM8,可以同时绘制72份南瓜属种质资源的指纹图谱。所有供试材料用5对多态性SRAP引物即可全部区别开来。研究表明,SRAP分子标记技术可成功地绘制南瓜属种质资源DNA指纹图谱。本研究对南瓜属种质资源鉴别、分子数据库构建及品种权保护具有较重要的意义。     

三倍体白杨杂种无性系指纹图谱的构建

目的:构建三倍体白杨杂种无性系指纹图谱,鉴定三倍体白杨杂种无性系。方法:分离纯化三倍体白杨杂种DNA模板,采用扩增片段长度多态性(AFLP)分子标记技术构建三倍体白杨杂种无性系指纹图谱。结果:从64对引物组合中筛选出M-CTA/E-CAG、M-CAC/E-CCA、M-ACT/E-CTC和M-CTT/E-CTG等4对多态性较高的引物组合,并应用该引物组合对21个三倍体白杨杂种无性系进行了AFLP分析,构建了21个三倍体白杨杂种无性系指纹图谱。结论:构建无性系指纹图谱是鉴别三倍体白杨杂种无性系的有效方法,能够有效鉴别21个三倍体白杨杂种无性系。本研究为品种鉴定及新品种权保护奠定了基础。     

遗传标记及其在作物品种鉴定中的应用

本文评述了用于作物品种鉴定的形态标记（morphological markers）、细胞标记（cytological markers）、生化标记（biochemical markers）、分子标记（molecular markers）的优缺点。重点评述了分子标记在作物品种鉴定中的应用。文中除对蛋白质电泳指纹图谱——同工酶和贮藏蛋白（包括醇溶性蛋白、清蛋白、谷蛋白、球蛋白等）电泳产生的指纹图谱的应用外,较详细地介绍了近年来DNA指纹图谱技术;包括限制片段长度多态性（restriction fragment length polymorphism,简称RFLP）、随机扩增多态性DNA (random amplified po lymorphic DNA,简称RAPD)、小卫星DNA（minisatellite DNA）、微卫星DNA（microsatellite DNA）,简单重复序列间扩增（intersimple sequence repeats,简称ISSR）,扩增片段长度多态性（amplified fragment length polymorphism,简称AFLP）以及CAPS (cleaved amplified polymorphic sequences)和SNPS (single nucleotide polymorphisms)对作物品种鉴定和新品种登记,品种纯度和真实性的检验以及品种间亲缘关系的探讨和在分类研究中的贡献等。     

甘薯质体遗传方式的DNA指纹图谱分析

用DNA指纹图谱分析了甘薯(Ipomoea batatas Lam.)徐薯18和AB78-1品系及它们的正反交子代叶绿体DNA,结果显示子代叶绿体DNA指纹均与母本相同,而未发现与父本或双亲相同的指纹图谱,因此在该杂交组合中质体遗传方式为母系遗传.这个结论与先前根据细胞学研究所推测的甘薯质体遗传方式不同,表明旋花科植物可能并不存在一个一致的父系或双亲质体传递模式.DNA指纹图谱分析用于质体遗传的研究尚未见报道,本文对其优越性进行了讨论.     

Use of SSR markers to complement tests of distinctiveness, uniformity, and stability (DUS) of pepper (Capsicum annuum L.) varieties

This study was carried out to assess the potential of SSR markers for variety identification by comparing SSR markers and morphological traits in tests of distinctiveness, uniformity, and stability (DUS) of pepper (Capsicum annuum L.) varieties. Twenty-seven SSR markers were polymorphic in 66 pepper varieties, revealing a total of 89 alleles. Average polymorphism information content (PIC) value was 0.529, ranging from 0.03 to 0.877. Cluster analysis of the band patterns separated the varieties into three groups corresponding to varietal types. Morphological trait-based clustering showed some degree of similarity to dendrogram topologies based on the SSR index. However, no significance correlation was found between the SSR and morphological data. SSR markers could be used to complement a DUS test of a candidate variety and to select complimentary varieties by pre-screening existing varieties in the context of protecting new varieties of pepper.     

SRAP标记与形态学标记在西瓜DUS测试中的比较

DUS（特异性、一致性和稳定性）测试是进行新品种申请的必要步骤。本文以28个不同的西瓜品种为研究对象,分别采用21对SRAP引物标记和54个用于DUS测试的形态学标记对其进行遗传多样性分析,其中SRAP引物在不同品种间的多态信息含量（PIC）在52.5%~89.2%之间,平均值为72.0%,计算得到的各材料间相似系数在0.92至0.99之间,而形态学标记统计得到各材料间相关系数在0.50到0.85之间。采用UPGMA法对所有材料进行聚类分析,SRAP分子标记聚类划分成四类,形态学标记将其划分为五类。对两种标记所得的结果进行相关性分析得出两者的相关系数为0.218,表明形态学标记和SRAP标记在这些材料上表现的相关性不是很高,但在品种鉴定和区别上SRAP标记表现出一定的优势,可以作为DUS测试的一种有益补充。     

Distinctness, uniformity and stability testing of tomato varieties in Poland

Testing for (DUS) in tomato varieties is a statutory requirement before varieties can be entered into the Register of Cultivars (National List) and/or granting of Plant Breeders’ Rights (PBR). The procedure and methods used for DUS testing of tomato varieties are outlined. In Poland all activities connected with plant variety testing and the maintaince of the Register of Cultivars and the Register of Plant Breeders’ Rights are provided by the Research Centre for Cultivar Testing (COBORU). The DUS testing procedures for Polish requirements are based on the Union for the Protection of New Varieties of Plants (UPOV) guidelines.     

六出花属DUS测试性状筛选与评价

以39份六出花品种为试材,对51个候选性状进行观测,比对不同测量方式对花相关数量性状的影响,并同时研究了同品种中外花被片与侧外花被片长度的相关性。结果表明:"花序:花序梗长度"和"花:花梗长度"不满足DUS测试性状要求,最后筛选出43个性状作为DUS测试性状。12个数量性状可分别划分为3~7级;不同观测方式对花相关数量性状具有影响,推荐DUS测试时采用随机观测方式;不同批次的同一品种观测结果部分性状上存在差异;多数品种的中外花被片长度与侧外花被片长度之间呈线性正相关;形态性状的聚类分析结果支持将"花:主色"作为分组性状。     

Testing uniformity of mutants of the Lathyrus sativus L. (grasspea) using Bennet's method

In this paper interest is focused on uniformity of individual mutants. Because before the registration of a new variety the criteria for DUS (distinctness, uniformity and stability) are checked, so you should also check whether the mutant meets them. The aim of created mutants, in many genetic experiments, is to expand variation of the initial material. This is achieved through use of the mutagen or combination of mutagens on different stages of plants growth. The experimenter should choose some of obtained mutants (for example these which showed better property as regards the same trait). Calculations were done on the basis of results of test with variety Krab of the Lathyrus sativus L. (grasspea) species and 17 mutants getting from this variety. Uniformity was checked with the use of the Bennett's method which is based on testing of equality of coefficients of variation. After analysis the mutants with codes K3, K37, K63 and K64 were detected as uniform at the same level as variety Krab.     

Testing uniformity of mutants of the <Emphasis Type="Italic">Lathyrus Sativus</Emphasis> L. (grasspea) using Bennet’s method

In this paper interest is focused on uniformity of individual mutants. Because before the registration of a new variety the criteria for DUS (distinctness, uniformity and stability) are checked, so you should also check whether the mutant meets them. The aim of created mutants, in many genetic experiments, is to expand variation of the initial material. This is achieved through use of the mutagen or combination of mutagens on different stages of plants growth. The experimenter should choose some of obtained mutants (for example these which showed better property as regards the same trait). Calculations were done on the basis of results of test with variety Krab of the Lathyrus sativus L. (grasspea) species and 17 mutants getting from this variety. Uniformity was checked with the use of the Bennett’s method which is based on testing of equality of coefficients of variation. After analysis the mutants with codes K3, K37, K63, and K64 were detected as uniform at the same level as variety Krab.     

Molecular determinants of dihydrouridine synthase activity

Dihydrouridine is one of the most abundant modified bases in tRNA. However, little is known concerning the biochemistry of dihydrouridine synthase (DUS) enzymes. To identify molecular determinants that are necessary for DUS activity, we have developed a DUS-complementation assay in Escherichia coli. Using this assay, we have identified amino-acid residues that are critical for the activity of YjbN, an E. coli DUS. We also show that the aq1598 gene product, a putative DUS from Aquifex aeolicus, catalyzes dihydrouridine formation, providing the first biochemical demonstration that A. aeolicus encodes an active DUS.