叶绿体是植物细胞中重要的细胞器，大部分叶绿体蛋白质都是由核基因组编码，在细胞质中合成分子量较大的前体蛋白，转运至叶绿体实施其功能。TOC33/TOC34是叶绿体上发现的一个外膜蛋白转运器构件蛋白，它与TOC159、TOC75和TOC64相互作用，构成了叶绿体外被膜上的一个蛋白转运器[1]目前已从豌豆(Pisumsa tivum)、拟南芥(Arabidopsis thaliana)、玉米(Zea mays)、小立碗藓(Physcomitrellapatens)、诸葛菜(Orychophroginus violaceus)和油菜(Brassica napus)克隆到TOC33或TOC34的CDNA或DNA的编码区。与其功能研究相比，TOC33基因的表达调控研究较少，该基因5’端调控区域的克隆及序列分析均未见报道。为此，在本实验室已经克隆到甘蓝型油菜TOC33基因编码区的基础上，采用单引物PCR方法[2]进行染色体步移，克隆出TOC33基因的启动子，为进一步研究TOC33基因的转录调控机制奠定基础。

用两个分别能在“蜀杂9号”父、母本之司稳定产生多态陆的随机引物S18和S31，对父、母本DNA样品进行RAPD扩增，将得到的特异标记片段S18750和S31550进行回收、克隆和测序。根据测序结果设计序列特异扩增(SCAR)引物SZ9SP1和SZPS92，将“蜀杂9号”的亲本RAH]标记转化为序列特异的SCAR标记。当用两对引物分别扩增杂种F1代单株总DNA时，能分别获得父、母本特异序列扩增带，表明获得的SCAR标记能可靠地用于“蜀杂9号”杂种纯度的检测。

A double low homozygous two-type sterile line ZY-2AB, its near-isogenie temporary maimainer line ZY-4, and the full sterile line ZY-3 derived fi'om the cross of ZY-2A with ZY-4, were developed to breed three-line dominant (MS hybrid in Brassica napus. Three yeats data showed that the ratios of sterile and frtile plants fit the expected 1:1 ratio in ZY-2AB, and the 1:0 ratio in ZY-3, respeetively. Some double low super hybrid combinations were screened out when the full sterile line ZY-3 was used as female par-ent, among which the combination 01Z24 increased 36.88 pereem more yield than eomrol and the combi-nation 98Z131 has been passed through the Siehuan Provincial Regional Test and registered in Siehuan Province. Two years data showed that the agronomic performance of the two-line hybrids derived from crosses of ZY-2A with re.storer lines versus that of the three-line hybrids produced by ZY-3 was not dirffe-ent for any of the nine most important agronomic characteristics, re.vealing that near-isogenie line of ho-mozygous two-type line was used as temporary maimainer could effectively avoid the disadvantage of three-way cross hybrid in breeding three-line dominant GMS hybrid.

用从100个10merS随机引物中筛选出的13个具有特异多态性引物对黑子南瓜种质资源遗传多态性进行了PAPD(Random amplified polymorphic DNA)分析。结果表明，PAPD对中国南瓜种、印度南瓜种、美洲南瓜种及黑子南瓜种的分类与传统分类学的结果相符。我国现有栽培黑子南瓜在DNA水平上的同源性极高，种质资源的遗传变异度较小，多样性水平较低，不如其它三个南瓜种多态性丰富。

According to the sequence of a 352bp fragment-M17 from the suppression subtractive hybridization library of the mutant line Cr3529 of Brass/ca napus, primers were designed and a 1000bp upstream fragrnet was cloned by 5'-RACE, which was joined with the original piece and conposed a 1343bp sequence. According to the 3' and 5' ends sequencess, a pair of primers were designed and the encoding region sequence of this unknown protein was amplified and cloned. Se-quence analyzing showed that it had 80.1% identity with an tmlmown-protein-encoding gene from Arabidopsis thaaliana. Swiss-model was used to predict the structure of the induced protein, and a potential transmembrane helix from site 46 Leu to 66 Phe was identified. GLOBE prediction of globularity showed that this protein possessed a compact globular do-main.