与黄瓜抗枯萎病基因连锁的RAPD标记

以黄瓜抗枯萎病亲本WIS2757和感枯萎病亲本津研2号及其F2分离群体为试材,采用分离群体分组分析法(BSA)进行了与黄瓜抗枯萎病基因连锁的分子标记研究。运用RAPD技术,利用780条RAPD引物对抗、感亲本进行筛选,其中有113条引物在两亲本之间表现多态性,但仅有引物S49在两组间多态性标记与亲本的多态性标记相同。经F2单株分析,引物S49扩增出的特异DNA片段与WIS2757抗黄瓜枯萎病基因连锁,遗传距离为14 cM。DNA标记条带大约为300 bp,定名为S49-300。     

Mutation Techniques for Gene Discovery and Crop Improvement

One of the most important breakthroughs in the history of genetics ）was the discovery that mutations can be artificially induced in organisms （van Harten, 1998）. Artificially induced mutations, by physical and chemical mutagens, have greatly advanced the understanding of genetics of higher organisms. Starting in the late 1960＇s, the International Atomic Energy Agency （IAEA） and the Food and Agriculture Organization （FAO） of the United Nations sponsored extensive research on mutation induction and their application to breeding of food and indnstrial crops that resulted in the introduction of new varieties of rice, wheat, barley, apples, citrus, sugar cane, banana, and others （more than 2 500 officially released new varieties are to be found in the FAO/IAEA Mutant Varieties Database） （hitp：//www-mvd.iaea.org/MVD/default.htm）. However, the usefulness of mutation techniques has been underappreciated in research communities, particularly during the last decade, when more and more researchers and breeders were rushing into molecular marker techniques and transgenic plants. In this paper, after a brief review of the past accomplishments of mutation induction and its application, we discuss the uniqueness of induced mutations in gene discovery and how to integrate induced mutants into functional genomics programs;     

TILLING and Ecotilling the Plant Kingdom

The modem crop scientist has a large amount of available nucleotide sequence information to identify genes of potential agronomic importance. Using reverse genetic approaches, specific genes can be disrupted, and hypotheses regarding gene function directly tested in vivo. Although a number of reverse genetic methods have been introduced, many are limited in application because they are organism-specific, expensive, transgenic, or only transiently disrupt gene function. However, traditional mutagenesis using chemical mutagens has been widely used as a forward genetics strategy to create many new crop plant varieties at relatively low cost. Mutagens such as ethyl methanesulphonate （EMS） cause stable point mutations and thus produce an allelic series of truncation and missense changes that can provide a range of phenotypes （Greene et al., 2003）. TILLING （targeting induced local lesions IN genomes） is a high-throughput reverse genetic strategy that combines traditional mutagenesis and SNP discovery methods （Colbert et al., 2001; McCallum et al., 2000）. To identify mutations, target regions of-l.5 kb are amplified with fluorescently labeled gene specific primers. Heteroduplexes are then formed between wild-type and mutant strands, mismatches are cleaved by incubation with a single-strand specific nuclease,     

大白菜抗根肿病基因位点CRa和CRb的分子标记鉴定

利用大白菜抗根肿病基因CRa和CRb分子标记(SC2930和KBr H129J18R)引物组,对78份大白菜材料进行抗根肿病分子标记鉴定。结果表明,在这78份材料中,有34份材料含有SC2930-T(CRa抗病标记)标记,其中杂合抗病位点材料17份,纯合抗病位点材料17份。有37份材料含有KBr H129J18R抗病标记,其中纯合抗病位点材料15份,杂合抗病位点材料22份。有20份材料不含有CRa和CRb所对应的抗病标记,23份材料含有2个抗病标记。该研究初步明确了78份参试大白菜材料所含抗根肿病基因CRa和CRb类型,为大白菜抗根肿病育种提供材料选择依据。     

八眉猪DQA基因编码区多态性及生物信息学分析

采用克隆测序的方法对八眉猪DQA基因编码区进行多态性分析,并利用生物信息学方法预测八眉猪DQA基因的理化特性及其编码产物的结构功能。结果显示:八眉猪DQA基因编码255个氨基酸,编码区序列有14个碱基变异,11个氨基酸变异,外显子2区域表现出丰富的多态性。DQA基因编码产物是一种不稳定的水溶性蛋白,二级结构为混合型,三级结构由α-螺旋、β-折叠、β-转角和无规则卷曲组成。蛋白质功能预测结果显示,信号转导、受体、胁迫应答、免疫应答和生长因子的几率较高。研究结果为八眉猪的抗病分子育种和经济性状相关基因筛选提供理论依据。     

转反义Trxs基因小麦籽粒蛋白组分的巯基含量与α-淀粉酶活性的关系

利用反义trxs基因特异引物，对扬麦5号转反义trxs基因株系00TY5的T₄和T₅代进行了PCR分子鉴定，并对其纯合株系籽粒蛋白质组分的巯基含量、α-淀粉酶活性及二者的相关性进行了测试。结果表明，00TY5的T₄和T₅代PCR分子检测均呈阳性。与非转基因植株相比，在籽粒成熟和后熟过程中，籽粒清蛋白巯基含量在开花后30 d至成熟后5 d平均降低33.2%，球蛋白巯基含量在开花后20 d至成熟后5 d平均降低42.5%，均达极显著水平(P<0.01)；醇溶蛋白和谷蛋白巯基含量有下降的趋势，但降低幅度较小；α-淀粉酶活性低，变异小，尤其在开花后35 d至成熟后10 d，平均比非转基因植株下降36.5%。各种蛋白质组分的巯基含量与α-淀粉酶的活性呈正相关关系，清蛋白巯基含量在成熟前与α-淀粉酶活性相关性达极显著水平(P<0.01)；球蛋白巯基含量在成熟前10 d至成熟后10 d达显著水平(P<0.05)；贮藏蛋白巯基含量与α-淀粉酶活性的相关性在各时段均较高。     

稻瘟病抗源筛选和抗病基因分析

本文介绍了抗源品种和育成品种的抗病性鉴定和筛选方法,指出多年多点鉴定、分菌系鉴定、混合菌系鉴定和特殊小种鉴定等多种方法前综合利用,能够准确地反映品种的抗病性,有效地筛选抗源品种和应用品种。应用这种鉴定和筛选方法已从云南粳型品种中选出一批可供抗病育种利用的新抗源。为了深入地了解品种的抗性,本文还介绍了抗病基因的推断方法及抗病基因分析方法。     

对小麦白粉病菌生理小种三个鉴别寄主的抗性基因分析

采用基因推导法分析了三个鉴别寄主的抗性基因：白免３号和肯贵阿１号含有Ｐｍ４ａ：小白冬麦的抗性受一对隐性基因的控制，其抗谱不同于所有供试单基因系的抗谱。     

玉米胚乳突变基因与互作对籽粒成份的影响 Ⅲ.su_1基因与sh_2、bt_2基因的互作效应及其利用价值

研究了su_1基因与sh_2,bt_2基因互作对玉米籽粒粒重及营养成份的影响,探讨了这些互作效应对玉米品质育种的利用价值。结果表明:su_1基因与sh_2,bt_2基因互作大幅度降低籽粒百粒重,增加可溶性糖、还原糖和蔗糖含量,降低淀粉含量,增加蛋白质及蛋白质组分中清蛋白、球蛋白和谷蛋白含量,降低醇溶蛋白含量。籽粒氨基酸组成中,天门冬氨酸、谷氨酸、丙氨酸、亮氨酸、赖氨酸、精氨酸等氨基酸受基因互作影响较大。籽粒发育过程中双突变体一般表现为:粒重早期与正常型及单突变体相差不多,但后期增长缓慢;淀粉、醇溶蛋白早期含量较低,后期增长幅度亦较小;蛋白质含量下降量较小;可溶性糖及还原糖早期含量较高,后期下降速度随时期或基因组合而异。研究表明,su_1基因与sh_2、bt_2基因互作可以进一步提高籽粒营养成份,对玉米品质改良具有特殊的利用价值,但提高粒重是这类基因互作效应应用的关键。     

利用RFLP／AFLP构建水稻褐飞虱抗性基因Bph1的连锁图

褐飞虱(Nilaparvata lugens Stal)是水稻最严重的害虫之一。已有人探索和利用天然抗性系统解决水稻生产中的褐飞虱问题。到目前为止，在本地籼稻品种和两个野生近缘种澳洲野稻和药用野生稻中鉴定到至少12个褐飞虱抗性主基因。