黑木耳激光诱变及营养缺陷型突变体筛选

利用 He- Ne激光和 YAG激光照射黑木耳担孢子诱发突变 ,共获得 14株营养缺陷型突变菌株 ,并用生长谱法和营养物逐个减少法对缺陷型类型进行了鉴定。实验结果表明 ,利用激光技术诱发黑木耳突变效果良好 ,其突变频率 (0 .0 5～ 0 .19)比传统的紫外线法明显提高。     

Isolation and mutation rate of the spontaneous PUR box and purR mutants

In our previous research in purine metabolism in Salmonella typhimurium, it was observed that the mutation frequency of the PUR box was one order of magnitude higher than that of purR under mutagenesis. In order to investigate further into this phenomenon, large amounts of independent PUR box and purR spontaneous mutants were isolated on lactose minimal medium by using a super-repressing mutant of purR (purRs). The mutational regions of 5 PUR box mutants and 4 purR mutants were cloned. Nucleotide sequence analysis showed that all the mutants had mutations at the expected sites. The comparison of the two types of mutations indicated that, although the purR gene was two orders of magnitude larger than the box). Thus, we concluded that high mutation frequency of the PUR box did not result from mutagenesis. Under spontaneous conditions, the mutation frequency of PUR box was also high. Some tentative explanations of this interesting phenomenon are given in this report.     

植物体细胞无性系变异育种

体细胞无性系变异是继花粉和花药培养之后的又一种实用化的细胞工程育种新方法，体细胞无性系变异是植物组织培养过程中出现的普遍现象，并且绝大多数变异可以遗传，在再生植株中能够找到在常规诱变育种和杂交育种中所观察到的各种变异或重组类型，80年代以来，我国在作物无性系变异育种方面取得了令人鼓舞的进展，已培育出了一大批优良新品种，本文着重阐述了植物体细胞无性系变异育种的概念和特点，引起无性系变异的遗传基础，无性系变异突变体的筛选和鉴定方法，并对目前体在植物体细胞无性系变异育种中存在的主要问题进行了讨论。     

小麦蜡质突变体Wx蛋白突变基因的分子鉴定

蜡质型小麦面粉对于改善面食品的加工品质、适口性、风味特点和延长货架寿命具有重要意义.淀粉粒结合型淀粉合酶I被称为蜡质蛋白(Waxy Protein,简称为Wx蛋白),是决定胚乳中直链/支链淀粉比例的关键酶.Wx基因发生突变并不影响产量等其它性状的表现.小麦三个Wx基因位点中Wx-D1的天然突变型遗传资源非常稀有.本实验对EMS诱导扬麦158的6个蜡质型突变体及其子代系进行了基于PCR的遗传多态性分析、单粒种子Wx蛋白SDS-PAGE电泳鉴定、PCR产物的克隆分析.利用自行设计的引物为5个突变体子代找到了STS标记;查明了两个突变体突变位点及其突变方式,是新的突变方式.     

激素对甘蓝型油菜(Brassica napus L)矮化突变体幼苗生长及内源GA3含量的影响

以甘蓝型油菜矮化突变体“DDF-1”为材料，研究了赤霉素（GA3）、生长素（IAA）和油菜素内酯（BR）3种激素对突变体幼苗的影响，并用石蜡切片观察其细胞形态，用酶联免疫吸附技术(ELISA) 检测下胚轴内源激素GA3含量.结果表明，外加一定浓度的GA3（7 mg/L）对矮化突变体“DDF-1”的下胚轴伸长作用显著，但不能使之恢复到野生型的长度；萌发早期，BR有明显的促进伸长作用，萌发后期效果不明显；突变体对IAA敏感性较弱.三种激素交互作用对矮秆下胚轴伸长的影响也不显著.施加外源GA3的矮化突变体“DD     

稻瘟病菌粗毒素对水稻幼胚愈伤组织的影响及抗性筛选

利用目前稻瘟病不同的优势小种菌株的液体培养提取的粗毒素对水稻幼胚进行处理,稻瘟病菌粗毒素提取液对水稻幼胚的愈伤的诱导和分化的有明显的抑制效应.随着毒素浓度的增加,水稻胚性愈伤获得率和分化能力显著降低.在诱导培养基中加入毒素对幼胚的筛选取效果较好,可以获得较多的胚性愈伤和较多的分化苗.培养基中毒素的浓度处理以30%效果好.水稻幼胚的抗性筛选过程中胚性愈伤获得和分化苗数较高,是很好的培养材料;而且不同的水稻品种产生的耐性不同,并可以同时筛选出抗稻瘟病的突变体.     

血红蛋白变异体(β15Trp→Leu)的功能与结构

目的 :用基因工程菌制造血红蛋白变异体 (β15Trp→Leu) ,并测定其生物功能 ,探讨血红蛋白分子结构与功能的关系 .方法 :通过对质粒pHE7的DNA分子操作用中性氨基酸Leu取代β15Trp ,利用层析分离技术等把血红蛋白变异体从基因工程菌中提取纯化出来 ,在确定其为目标蛋白质后进行稳定性、亲氧力与协同变构效应能力的测定 ,并对变异体生物功能特性进行分子结构的分析 .结果 :血红蛋白变异体 (β15Trp→Leu)结构稳定 ,与正常血红蛋白比较 ,亲氧力增强 ,协同变构效应能力下降 .结论 :β15Trp在血红蛋白分子结构中相当重要 ,由其与对应的氨基酸所形成的氢键与蛋白分子的生物功能关系密切 ,该氢键的失去使血红蛋白改变正常功能 ,亲氧力增强 ,协同变构效应能力下降 .     

小麦耐盐突变体及其亲本在生育中后期生理指标的比较研究

对小麦耐盐突变体H870 6 - 34 ,RH870 6 - 49及其亲本濮农 36 6 5 ,百农 30 39在生育中后期非盐胁迫下的SOD活性进行了比较研究。观察其在不同生育时期不同材料间的变化情况发现 ,在小麦生育中后期 ,耐盐性较好的材料RH870 6 - 49SOD活性相对较高 ,细胞质膜透性相对较低。而耐盐性较差的材料H870 6 - 34的SOD活性相对较低 ,细胞质膜透性相对较高     

叶绿素缺乏大麦突变体叶绿体结构功能及生化特性的研究

突变大麦类囊体膜吸收光谱的蓝区峰值明显高于野生型;A683/A652比值较高;CD谱信号较弱,这说明其捕光色素蛋白复合物发生了变化。SDS-PAGE结果表明,突变大麦的Rubisco的水平与野生型一致,但在类囊体膜上分子量为24-30kD的LHCⅡ蛋白组分减少。     

Construction and characterization of partiallyntrC-deleted mutants inAlcaligenes faecalis

To study the effect of ntrC gene product on the expression and regulation of other important nitrogen-fixing genes in Alcaligenes faecalis, partially ntrC-deleted mutants of A. faecalis have been generated. To start with, the ntrC gene of A. faecalis was cloned into a suicide plasmid pSUP202 to create a recombinant plasmid pSUM1. The ntrC gene in pSUM1 was then replaced by a lacZ-Km^r fragment resulted in the generation of a plasmid pSUM2. The lacZ fragment in pSUM2 was further removed and a plasmid pSUM3 produced. As a second step, the plasmid pSUM2 or pSUM3 was introduced into the wild type of A. faecalis A1501 by conjugation and two partially ntrC-deleted mutants A15CM1 (ntrC∷lacZ) and A15CM2 (ntrC ^- ) were obtained. To understand the regulatory effect of the NtrC on the expression of nifH and nifA, a nifH-lacZ gene or a nifH-lacZ gene was introduced into the ntrC^- mutant by conjugation. The results indicated that: (ⅰ) although the ntrC^-mutant was nif ⁺ , its nitrogen fixation activity was only 20% that of the wild type; (ⅱ) the ntrC^- mutant failed to grow on the medium containing nitrate as a sole nitrogen source; (ⅲ) the regulation of ntrC gene expression did not require its own product; (ⅳ) the expression of nifH in A . faecalis was positively regulated by the ntrC. Deletion of the ntrC resulted in the reduction of nifH expression or even totally inactivated nitrogen fixation; (ⅴ) there was no obvious influence on the expression of nifA in A. faecalis if the ntrC gene was deleted.