DNA分子标记技术在水产养殖中的研究应用

分子标记是继形态标记、细胞标记和生化标记之后发展起来的一种比较理想的遗传标记技术。本文综述了DNA分子标记的类型,基本原理和特点,以及分子标记辅助育苗在水产养殖方面取得的可喜进展。     

烟叶果胶糖醛酸结构的固态核磁共振波谱表征

烟叶果胶不同糖醛酸结构的含量分布,对其生理功能和产品品质有着重要影响。为表征烟叶果胶糖醛酸结构,采用固态¹³C交叉极化/魔角旋转核磁共振波谱技术(¹³C CP/MAS NMR),以二甲基硅橡胶管为内标物质,建立烤烟烟叶中果胶含量的定量分析方法。选择NMR图谱中果胶C-6区域的波谱峰,应用PeakFit软件进行去卷积分峰处理,实现烟草样品果胶不同糖醛酸结构,即果胶酸(-COOH)、甲酯化果胶(-COOCH₃)、果胶酸钙盐(-COO^-)等定性和含量分布的分析表征。应用固态核磁共振波谱分析表征了不同部位9种烟叶样品中的果胶含量及其糖醛酸结构。结果表明,烟叶果胶含量随着烟叶部位上升而逐渐增高。样品烟叶中甲酯化果胶的含量分布均小于50%,属低甲酯化果胶。对比中部烟叶,上部叶的甲酯化果胶含量偏高,下部叶偏低,而上部叶的果胶酸钙盐含量较低,下部叶较高。结果为烟草果胶大分子结构与性能的关系研究,提供准确、简便的技术方法。     

棉花QTL定位及MAS育种进展与展望

近年来,随着现代分子生物学及统计分析方法的快速发展,育种工作者已定位了许多作物的重要性状QTLs.同时,基于QTL定位的标记辅助选择(MAS),已成为当前作物分子育种研究的热点.综述了近年来棉花重要性状包括产量、纤维品质、抗性、形态、生理、早熟性等QTL定位最新进展,以及标记辅助选择在一些性状上的初步应用,并对其发展前景进行展望.     

猪育种标记辅助选择的经济效益研究

为了研究在育种群中利用MAS对猪育种中所需选择的5个性状即NBA、BF、FCR、ADG、IMF的经济效益,本研究采用3个不同规模的基础群:母猪分别为100、200、300头,公猪都为10头,利用LE、LD和DR等3种类型标记对5个性状分别进行MAS闭锁繁育选择5个世代,基础群个体间无亲缘关系;估计育种值采用的模型为SBLUP(标准BLUP)模型(以此育种值为对照)、MBLUP混合模型(使用LE和LD标记时)和QBLUP模型(利用DR标记时).由各性状利用MAS获得的额外遗传进展计算MAS带来的经济效益.研究结果表明,利用DR标记获得的经济效益高;3个规模不同育种群利用DR标记进行MAS时,300头母猪群体获得的经济效益最高;尽管利用LE标记比利用LD标记获得更多的额外遗传进展,但还是以利用LD标记获得的经济效益更高.     

分子标记辅助选择Xa23基因培育杂交稻抗白叶枯病恢复系

水稻抗白叶枯病新基因Xa23抗谱广、抗性强,被定位在第11染色体上。以携带抗白叶枯病基因Xa23的抗病品系CBB23为抗源,以优良杂交稻亲本9311和1826为受体材料,采用杂交和复交,在分离群体中利用与Xa23紧密连锁的EST标记C189进行分子标记辅助选择,通过苗期分子标记检测和成株期农艺性状选择,获得160份目标基因纯合且农艺性状稳定的株系。使用鉴别菌系P6,采用人工剪叶接种法,在苗期和孕穗期对21份重点株系进行了抗性鉴定,所有株系在苗期和孕穗期都表现抗病。同时对3个不育系与其中5个株系配制的15个杂交组合进行了抗性鉴定,所有组合在苗期表现抗或中抗,在孕穗期表现抗。对其中6个杂交组合进行了品比试验,2个组合产量略低于对照两优培九,其余4个组合的产量均高于两优培九,用于测配的3个株系C6201、C6271和C6351有望作为杂交稻恢复系在生产中应用。研究表明在育种进程中利用与Xa23基因紧密连锁的分子标记C189开展抗白叶枯病分子标记辅助育种是一种有效的途径。     

Aeromonas hydrophila and Aeromonas veronii cause motile Aeromonas septicaemia in the cultured Chinese sucker,Myxocyprinus asiaticus

In August 2017, a serious disease causing high mortality occurred in a Myxocyprinus asiaticus aquaculture farm. According to necropsy findings, bacteriology experiments and phylogenetic analysis based on 16S rRNA, cpn60, gyrB and rpoB genes and concatenated alignment sequences (cpn60, gyrB and rpoB genes), two isolates, that is, BBAh1 and BBAv1, were identified as Aeromonas hydrophila and Aeromonas veronii respectively. Artificial infection experiments were carried out, showing that the BBAh1 and BBAv1 strains can cause similar symptoms and have LD50 values of 1.93×10⁵ and 8.77×10⁵ cfu/g respectively. In addition, some virulent genes coding for AerA, Alt, Ast, AscV, AexT, AspA, HlyA, OmpA, Lip and FlaA were detected in the two strains. Furthermore, BBAh1 and BBAv1 showed the same sensitivities to 28 antibiotics, some of which, such as cefotaxime, aztreonam, ceftriaxone and tetracycline, may be used to control the disease. However, the strains were also resistant to many antibiotics. These results provide a scientific reference for the diagnosis, prevention and treatment of motile Aeromonas septicaemia caused by A. hydrophila or A. veronii in cultured Chinese sucker.     

运用分子标记辅助选择技术改良9311白叶枯病抗性的研究

以9311为轮回亲本，以IRBB21为抗白叶枯病供体亲本，通过杂交和三次回交以及两次自交，结合农艺性状的选择，建立起具44个株系的BC3F3群体。分别用连锁标记pTA248和基因内STS标记MXA21对抗性基因Xa21进行前景选择。用均匀分布在水稻全基因组的248个SSR标记，进行两亲本之间的多态性分析，获得76个具多态性的SSR标记，用于背景选择。结果选到基因型背景回复到轮回亲本的73．68％——88．16％，且Xa21基因纯合的株系4个，分别为M071、M082、M086和M087。以Xa21的鉴别菌系菲律宾6号小种PX099接种鉴定表明，9311为感(S)，IRBB21为抗(R)，中选株系M071和M082的抗性与IRBB21相同，为抗(R)级，M086和：M087抗性略低于IRBB21，为抗到中抗(R／MR)，这两个株系内单株之间抗性有差异。农艺性状考察结果显示，中选株系与轮回亲本9311相似。以中选株系M086与培矮64S配制的Fl和原组合两优培九(培矮64S／9311)进行比较，对白叶枯病的抗性有了明显提高，综合农艺性状相似。论文就白叶枯病抗性基因的综合利用以及育种方法进行了讨论。     

华南抗稻瘿蚊分子标记辅助育种

亚洲稻瘿蚊(Orseolia oryzae Wood-Mason)是华南的主要水稻害虫.选育抗虫品种是最有效的生态控制方法.本文综述1998-2006年抗性育种的进展.用AFLP方法对从中国广东省7个地点采集的4个生物型的DAN指纹进行分析;在对用RAPD和SSR技术分别对抗中国4个稻瘿蚊生物型的基因Gm6精细定位基础上,用与Gm6紧密连锁的STS和SSR标记开展分子标记辅助育种,创造了一批抗稻瘿蚊的新种质,包括育成了6个栽培稻和6个二系杂交稻和1个三系杂交稻并在农户试种,在中国广东成功地建立了分子标记辅助选育抗稻瘿蚊品种的技术体系.     

STS markers for powdery mildew resistance gene Pm6 in wheat

The powdery mildew resistance gene Pm6, transferred to common wheat from the tetraploid Triticum timopheevii, is effective in most epidemic areas for powdery mildew in China. RFLP probe BCD135 was previously associated with Pm6. In the present research, four STS primers (NAU/STS_BCD135-1, NAU/STS_BCD135-2, STS003 and STS004) were designed from the sequence data of BCD135. These primers were used for PCR amplification using the genomic DNA of resistant near-isogenic lines with Pm6 and their recurrent parent, cv. Prins. No polymorphic product was observed using primers STS003 and STS004; however, primers NAU/STS_BCD135-1 and NAU/STS_BCD135-2 amplified two and one bands, respectively, polymorphic between the resistant near-isogenic-lines and Prins. The two primers were then used to amplify the F₂ population from the cross IGV1-465 (FAO163b/7*Prins) × Prins. The amplification and the powdery mildew resistance identification data were analyzed using the software Mapmaker 3.0. The results indicated that both NAU/STS_BCD135-1 and NAU/STS_BCD135-2 were closely linked to Pm6 with a genetic distance of 0.8 cM. A total of 175 commercial varieties without Pm6 from different ecological areas of China were tested using marker NAU/STS_BCD135-2 and none of them amplified the 230 bp-specific band. This marker thus has high practicability and can be used in MAS of Pm6 in wheat breeding programs for powdery mildew resistance. Jianhui Ji and Bi Qin contributed equally to this work.