水稻PCD相关基因的定位研究

植物中的细胞程序化死亡(PCD)在植物的发育、抗病及植物与环境互作等过程中发挥着极其重要的作用.本研究以我国学者发现的特殊的水稻细胞死亡控制突变体3037(M)为材料, 采用微卫星DNA标记 (共计159个微卫星引物) 技术对突变体性状进行基因定位研究.结果表明, 该突变体PCD基因位于水稻的第8染色体和第12染色体,为下一步精细定     

棉花表型性状基因的SSR标记定位

本文以两个陆地棉多标记基因系T582和T586,以及杂交获得的F_1、F_2及F_3代作为试验材料,利用11对SSR引物对F_2群体的120个单株的DNA样品进行多态性分析,并利用F_2和F_3群体对F_2群体对应单株的13个表型性状进行基因型的判定,结果得到了3个与表型性状基因连锁的SSR标记,分别是红茎基因(R_1)与J178连锁、遗传距离为24.9cM,簇生铃基因(CL_1)与J236连锁,遗传距离为46.0cM,茸毛基因(T_1)与J252连锁,遗传距离为28.5cM,其中R_1、CL_1、J178和J236在同一连锁群上。红茎和植株茸毛是具有抗虫性能的形态性状,用SSR标记这些性状将有助于提高育种家的育种效率。     

水稻早世代稳定特异性的研究

利用9个具有早世代稳定遗传特性的水稻品系和7个栽培稻作为主体亲本杂交配组，在130个组合F2群体963个株系中，发现28个组合中出现73个稳定株系。遗传分析表明水稻早世代稳定既不是质量性状遗传也不是数量性状遗传，是按株群分离的遗传方式，出现稳定株系的组合F1群体发生了分离，F2群体中既有性状不分离的稳定株系又有孟德尔     

转Bt-CpTI-GNA基因棉花的研究

采用农杆菌介导法将外源三价抗虫基因(Bt-CpTI-GNA)导入常规棉花品种中,获得转基因再生株,分子检测表明外源基因已在棉花体内表达,并遗传给后代材料。PCR分子检测与转化的标记基因和外源目的基因抗性三者极有规律性。其所携带的基因在转基因棉花中有分离现象,这可能是外源基因整合到受体棉株体内“基因沉默”而引起所致。     

Genetic mapping of AFLP markers in Japanese bunching onion (<Emphasis Type="Italic">Allium fistulosum</Emphasis>)

Summary The first genetic linkage map of Japanese bunching onion (Allium fistulosum) based primarily on AFLP markers was constructed using reciprocally backcrossed progenies. They were 120 plants each of (P₁)BC₁ and (P₂)BC₁ populations derived from a cross between single plants of two inbred lines: D1s-15s-22 (P₁) and J1s-14s-20 (P₂). Based on the (P₂)BC₁ population, a linkage map of P₁ was constructed. It comprises 164 markers – 149 amplified fragment length polymorphisms (AFLPs), 2 cleaved amplified polymorphic sequences (CAPSs), and 12 simple sequence repeats (SSRs) from Japanese bunching onion, and 1 SSR from bulb onion (A. cepa) – on 15 linkage groups covering 947 centiMorgans (cM). The linkage map of P₂ was constructed with the (P₁)BC₁ population and composed of 120 loci – 105 AFLPs, 1 CAPS, and 13 SSRs developed from Japanese bunching onion and 1 SSR from bulb onion – on 14 linkage groups covering 775 cM. Both maps were not saturated but were considered to cover the majority of the genome. Nine linkage groups in P₂ map were connected with their counterparts in P₁ map using co-dominant anchor markers, 13 SSRs and 1 CAPS.     

Construction and characterization of a BAC library of soybean

We have constructed a soybean (Glycine max (L.) Merrill) bacterial artificial chromosome (BAC) library from green leaf protoplasts of the cultivar, Misuzudaizu. The library contains 53,760 clones with an average insert size of 116 kb. About 2.9% chloroplast DNA origin was revealed by PCR and colony hybridization. Apart from 2.8% clones having no insert, this library represents 5.2 genome equivalents. With this genome coverage, the probability of having any DNA sequence represented in the library is higher than 99.5%. Three-dimensional pools of the BAC library in combination with the use of a high efficiency genome scanning (HEGS) electrophoresis facilitate rapid and efficient PCR-based screening. An average of five positive clones were identified after screening the BAC library with SSR and STS markers. BAC-end walking was performed for three SSR associated BACs. This library will provide a good resource for positional cloning of agronomically and biologically important QTL genes that Misuzudaizu possesses.     

SCAR marker tagged to the alien leaf rust resistance gene Lr19 uniquely marking the Agropyron elongatum-derived gene Lr24 in wheat: a revision

A sequence characterized amplified region (SCAR) marker tagged to an Agropyron elongatum‐derived leaf rust resistance (Lr) gene Lr19 was validated on 18 known alien Lr gener in near‐isogenic lines (NILs) in the variety ‘Thatcher’, along with three wheat cultivers carrying Lr24 and two carrying Lr19. The marker was expressed only in the Lr24 lines confirming that the marker tagged the geneLr24. The monomorphic expression of the SCAR marker in 10NIL pairs for Lr19 and Lr24 revealed that each NIL pair possessed the same gene, Lr24. The donor parents used in the NIL pairs for Lr19 (‘Sunstar*6/C80‐1′) and Lr24 (‘TR380‐14*7/3Ag#14′) amplified the same fragment. Nonsegregation for leaf rust in the F₂ population of the cross between the above donor parents confirmed the presence of the same gene in the two parents. Apparently, a genuine parent stock of ‘Sunstar*6/C80‐1’ was not involved in the development of the NIL pairs for Lr19 due to an improper maintence bredding protocol either at source or destination which went undetected in the absence of signs of virulence for either gene in the region.     

Development of a sequence-specific PCR marker linked to the Ku gene which removes the vernalization requirement in narrow-leafed lupin

Wild types of Lupinus angustifolius require vernalization to promote flowering. Modern domesticated cultivars carry the early-flowering gene Ku which removes this requirement. A microsatellite-anchored fragment length polymorphism marker was identified as co-segregating with the Ku gene in a recombinant inbred line (RIL) population derived from a domesticated × wild-type cross. DNA sequencing showed that the marker contained a 7 bp insertion/deletion polymorphism, as well as a single nucleotide polymorphism. A pair of sequence-specific primers was designed and successfully converted the size polymorphism into a simple polymerase chain reaction based co-dominant marker. This marker is closely linked to the Ku gene, as it co-segregates with the Ku phenotyping in a population consisting of 106 RILs.     

Variation of the genomic proportion of the recurrent parent in BC1 and its relation to yield performance in sorghum (Sorghum bicolor) breeding for low-input conditions

In order to define the variation of the genomic proportion of the recurrent parent [G(RP)] and its relation to yield, G(RP) of individual BC₁ plants of two sorghum populations composed of a high‐yielding cultivar as recurrent parent (RP) and a donor with superior drought resistance or grain quality, respectively, was estimated using AFLPs and SSRs. G(RP) in BC₁ ranged from 0.53 to 0.95 and averaged to 0.76 in the population (NP4453 × ‘SV‐2’) × ‘SV‐2’. G(RP) varied between 0.60 and 0.86 and averaged to 0.74 in the BC₁ of (ICV‐219 × ‘SV‐2’) × ‘SV‐2’. Results show that plants with a G(RP) equivalent to BC₂ (0.875) or BC₃ (0.938), respectively, can be selected from BC₁. Yield performance of BC₁S₁ families was tested in field trials carried out in South Africa. The correlation between yield and G(RP) in BC₁ was low. Selection according to G(RP) did not result in an effective preselection for yield.     

利用若干单株混合提取DNA方法进行玉米群体SSR的分析

以金皇后玉米群体90个单株：DNA和6个自交系DNA为供试材料，利用15对SSR引物，比较了同一群体4种DNA样品处理(单株DNA样品、3个单株混合DNA样品、10个单株混合DNA样品、15个单株混合DNA样品)的SSR遗传多态性。结果表明，利用单株DNA样品能获得等位基因数、基因频率、基因型杂合度等较全面的遗传信息，适宜进行单个或少量群体的遗传结构研究，但试验工作量大；利用混合DNA样品所能获得的遗传信息有所减少，比较4种DNA样品处理的检测结果发现，15个单株混合样品的检测结果误差较大，而采用3～10个单株DNA混合的样品基本可以反映出一个群体的等位基因数目和SSR带型的差异，由于试验工作量大大减少，可以应用于较多玉米群体间遗传差异的比较。