Construction and genetic analysis of mutator insertion mutant population in maize

A total of 26718 M1 plants were ob- tained by crossing the active mutator transposon donor parents (Q105, WW51, 115F, V26-2 and 919J) with the recipient parents (Hz85,W328 with Bz gene and S-Mo17Rf3Rf3). The phenotypes of M1 plants were observed in the field. M1 plants were self-pollinated to develop the mutator insertion-mutagenized M2 seeds. The transposition frequency of the mutator in the genome was calculated based on the spotted aleurone phenotype of the M2 seeds. The results showed that: (1) the mutation frequency of M1 phe- notypes in the field was 0.07 in the population of W328×Mu; (2) the mutation frequency of spotted aleurone seeds on the M2 ears was 0.122 in the population of W328×Mu; (3) five S-cytoplasm male-sterile plants were found among 22500 M1 plants of S-Mo17Rf3Rf3×Mu, with the transposition frequency about 2.2×10?4 per locus. 99 flanking se- quences of mutator transposition were amplified by the modified MuTAIL-PCR, and 59 non-redundant sequences with length around 400 bp were obtained. After bioinformatic analysis, 27 sequences of them could be annotated, using non-redundant nucleotide database of maize, rice, and Arabidopsis. 36 se- quences of them were located on the genetic map of maize by comparative genomics, and several flank- ing sequences of mutator insertion were mapped on the single marker locus. Hotspot sequences of mu- tator transposition were revealed by comparing the homologies between the 9-bp target site duplication of the mutator insertion. The putative functions of 8 flanking sequences of mutator transposition had identity with the functions of their correspondingmarker. The constructed mutator insertion mutant population in maize will facilitate the new gene discovery and functional genomics study in maize.     

高效液相色谱法测定谷子幼苗中的GA_1和GA_3

建立了同时测定谷子幼苗中赤霉素Ａ１（ＧＡ１）和赤霉素Ａ３（ＧＡ３）的高效液相色谱（ＨＰＬＣ）方法。采用ＯＤＳ Ｃ１８色谱柱,以磷酸盐缓冲液和甲醇为流动相,ＧＡ１和ＧＡ３在１０ｍｉｎ内获得良好分离。利用该方法对谷子的两个正常栽培品种及其矮秆突变体的幼苗进行了测定。测定结果表明,谷子幼苗中同时含有ＧＡ１和ＧＡ３两种活性赤霉素,它们各自的质量比是影响谷子株高的重要因素。     

紫花苜蓿抗赛性突变体的筛选

首先由紫花苜蓿叶片诱导产生愈伤组织，然后用ＥＭＳ对愈伤组织进行了处理，并进行了低温筛选，获得抗寒性突为体。并用生理生化方法对愈伤组织突变体的抗寒性进行了鉴定。     

转座子诱变甘蓝黑腐病菌所获胞外多糖突变体的类型

据对转转子Ｔｎ５ｇｕｓＡ５诱变甘蓝黑腐病民获得３９株胞外多糖突变休的３种胞外酶活性和在含２％葡萄ＮＹＧＡ培养基上的菌落形态的检测结果，将突变株分成５种类型。Ｓｏｕｔｈｅｒｎ杂交结果表明它们的突变位于基因组中的８个不同的位置，其中１个位于已鉴定的ｒｐｆ调控基因簇。     

水稻光敏核不育突变体与其原种的比较研究

本文对水稻光敏核不育突变体农垦５８Ｓ与其原种农垦５８进行了比较研究。结果表明，光敏核不育突变体农垦５８Ｓ与其原种农垦５８在各个生育期都存在明显的差异。农垦５８Ｓ表现出苗期株高、根长、叶绿素含量比农垦５８显著提高；生殖生长期，农垦５８Ｓ对光周期反应敏感，冠层叶片长度、株高、茎杆重和结实率在长日照或短日照处理下存在显著差异，农垦５８在生殖生长期对光周期反应不敏感。根据以上实验结果和已有的文献报道得出结论：水稻光敏核不育突变体农垦５８Ｓ不仅仅是雄性育性受育性转换期光周期调控这一个性状的突变，而且是同时伴随着多种性状的突变。     

坛紫菜色素突变体光合作用效率的比较研究

以野生型和5种不同色泽的坛紫菜色素突变体为研究对象,通过水下饱和脉冲荧光仪（DIVING-PAM）测定了不同色素突变体的光合作用参数,并测定了相应品系坛紫菜叶状体的4种主要光合色素（叶绿素（Chl.a）、藻红蛋白（PE）、藻蓝蛋白（PC）和别藻蓝蛋白（APC））的质量比。结果表明:坛紫菜不同色素突变体的光化学最大量子效率（Fv/Fm）、有效荧光产率（Y(Ⅱ)）、光化学淬灭系数（QP）和非光化学淬灭系数（QN)均表现出不同程度的差异,且没有明显的规律性,其中坛紫菜各品系藻体的Fv/Fm值均介于0.65~0.75之间,而Y(Ⅱ)值则随着光照强度的逐渐增强呈现为快速下降的趋势。光合作用参数和光合色素质量比的相关性分析结果则表明坛紫菜Fv/Fm值与各色素蛋白的质量比没有相关关系,却与w(Chl.a)/w(PC)和w(APC)/w(PC)极显著正相关;而Y(Ⅱ)除与w(Chl.a)/w(PC)和w(APC)/w(PC)极显著正相关外,还与w(PC)、w(APC)和w(Chl.a)极显著负相关。     

构建突变基因文库的定向进化研究进展

1.介绍定向分子进化是在实验室条件下模拟自然选择过程,改变原有蛋白的氨基酸序列,以期找寻预期性能的突变蛋白。定向分子进化实验的成功与否取决于制备的突变体库的质量和简捷、高效的筛选方法。随着体外定向进化分子改造新策略的不断涌现,以及相关领域如生物信息学、功能基因组学、功能蛋白质组学等的更新和完善,各种基因改组技术的应用正受到足够的重视。其中,酶工程的定向进化已为基因