Ploidy screening of anthurium (Anthurium andreanum Linden ex André) regenerants derived from anther culture

Anther culture was successfully developed in Anthurium andreanum Linden ex André cv. ‘Tropical’, but resulted in variation in morphology and growth response of regenerants. Ploidy levels varied in different morphological variants. Finding a convenient, rapid, reliable, practical and indirect method to screen and determine anthurium ploidy level is important not only for anthurium, but for ornamentals in general. Regenerants derived from anther culture showing three different ploidy levels (haploids, diploids, triploids) were used in this study. Five indirect methods were used to assess chromosome number: chloroplast number in stomatal guard cells (M1), stomatal length and width ratio (M2), stomatal density (M3), ratio of length and width of leaves (M4), and microspore number per anther (M5). These were compared to chromosome counting as the direct and control method (M6). Through simple regression correlation analysis, when compared to M6, M1 was the most convenient and reliable indirect method to determine the ploidy level. This method was highly and significantly positively correlated to anthurium ploidy level (r = 0.945; p < 0.01). This method could also be applied much faster than the conventional chromosome method.     

控制叶绿体运动光受体研究进展

综述了近年来有关叶绿体运动光受体研究的新进展。     

珍珠玉绿萝不同颜色叶片叶绿体色素含量差异

为揭示珍珠玉绿萝的斑叶呈色原因,以扦插繁殖的珍珠玉绿萝为材料,对其叶片不同颜色部分的叶绿体色素含量进行了测定分析。结果表明:叶片绿色部分的各种叶绿体色素含量及比值均远高于黄白部分,绿色部分与黄白部分的叶绿素(Chl)/类胡萝卜素(Caro)差异远大于其叶绿素a(Chla)/叶绿素b(Chlb)差异。因此,珍珠玉绿萝叶片中叶绿体色素的形成受抑导致黄白部分产生,尤其是Chla。     

基于叶绿体序列分析紫茎泽兰在菊科的系统发育

为研究紫茎泽兰(Eupatorium adenophorum Spreng)在菊科的系统发育,利用通用引物对紫茎泽兰叶绿体DNA的rpl16内含子及psbA-trnH与trnL-F两个基因间隔区进行PCR扩增并测序,将其序列分别与GenBank中部分菊科(Asteraceae)植物的基因序列进行比对分析。应用Mega 4.0软件进行分子遗传进化分析,采用邻接法构建系统进化树。结果表明,菊科植物在系统分类上呈现出两大类群,滨菊、野菊构成一个大类,和紫茎泽兰关系较远;紫茎泽兰和毛叶泽兰、圆叶泽兰、线叶泽兰等泽兰属植物及油菊的关系最近,与向日葵和银胶菊次之,与莴苣、滨菊、野菊关系最远,表明紫茎泽兰确实隶属于泽兰属;序列组合(rpl16+ps-bA-trnH+trnL-F)长度变异范围为2 140~2 203 bp,排序后的比对长度为2 399 bp,有381个变异位点,190个信息位点,为紫茎泽兰的系统进化提供了一定的信息。     

小麦叶绿体DNA提取方法研究

以成熟的中国春小麦叶片为材料,采用改进的高盐-低pH方法提取叶绿体DNA,经琼脂糖凝胶电泳和PCR扩增,结果表明:所提取的小麦叶绿体DNA纯度高(OD260/280=1.89,OD260/230=2.23),适于PCR反应、基因扩增和酶切鉴定等分子操作,为深入研究小麦叶绿体基因组奠定了基础。     

水稻“斑马叶”突变体B411叶绿体超微结构的观察

采用透射电镜技术,对水稻突变体B411"斑马叶"出现过程中叶片的叶绿体超微结构进行了研究。结果表明,在叶片包于叶鞘内的心叶期,叶片绿区叶绿体呈纺锤形,类囊体片层纵向整齐排列;黄区叶绿体呈不规则椭圆形,类囊体片层排列不规则。在叶片抽出叶鞘1d的嫩叶期,叶片绿区叶绿体类囊体片层增多并垛叠形成基粒结构,它们沿长轴整齐排列;黄区叶绿体类囊体膜断裂,片断化。在成熟期,叶片绿区叶绿体类囊体片层变得丰富和形成许多基粒结构;叶片黄区叶绿体内部结构严重解体,整个叶绿体呈高电子密度的囊泡状结构。在复绿期,叶片黄区逐渐变成绿色,叶绿体结构恢复正常,类囊体膜系统重建,且有序地沿叶绿体长轴方向排列,在基质中形成淀粉粒,表明其光合能力恢复。     

蝴蝶兰叶绿体基因组密码子使用的相关分析

通过对蝴蝶兰叶绿体基因组全序列密码子的研究,利用Codonw软件进行统计分析,得出密码子第三位置GC含量为31.41%,明显低于第一位置和第二位置的GC含量,说明密码子第三位置富含AT。从对应分析中得出其叶绿体密码子偏好性与其基因表达水平密切相关,且受到突变和选择压力的共同影响。将高表达优越密码子分析法和高频密码子分析法相结合,确定了20个蝴蝶兰叶绿体的最优密码子。上述结果对今后兰花的分子系统进化和高表达转基因研究具有重要意义。     

枇杷试管苗限制生长保存过程中叶肉细胞超微结构变化的研究

本试验以早钟6号枇杷试管苗为材料,将其分别保存在MS+5 mg/L PP333培养基上和MS培养基(对照)上,采用超薄切片透射电镜对保存的试管苗成熟叶片叶肉细胞进行观察。结果表明:与对照相比,添加5 mg/L的PP333,保存6个月时,明显推迟了叶绿体的变形、移动、叶绿体和线粒体的镶嵌及叶绿体的解体;保存9个月时,叶肉细胞叶绿体结构完整,少数出现解体,淀粉粒明显增大、增多,线粒体和细胞核结构正常。     

The distribution of normal and male-sterile cytoplasms in Chinese sugar-beet germplasm

Forty-two Chinese sugar-beet breeding lines were evaluated for the presence of normal and male-sterile (Owen) cytoplasms using polymorphisms in the chloroplast petG-psbE region as well as in the mitochondrial minisatellite loci. The polymorphisms detected allowed the distinction of three cytoplasm types over the whole sample, one being associated with Owen cytoplasm, a second with the maintainer inbred ‘TK-81mm-O’-type cytoplasm (termed normal-1 cytoplasm) and a third with another maintainer inbred ‘NK-310mm-O’-type cytoplasm (normal-2 cytoplasm). Western blot analysis was carried out to confirm that expression of the male-sterility-associated protein (preSATP6) occurred in plants with Owen cytoplasm but not in plants with either normal-1 or normal-2 cytoplasm. Of the 42 breeding lines examined, 14 had exclusively normal (normal-1 and/or normal-2) cytoplasm and 11 had only Owen cytoplasm. The remaining 17 lines possessed both normal and Owen cytoplasms, and noticeably, some of these 17 lines have been expected to become the source of superior maintainer lines. The results thus show that molecular identification of the cytoplasm is required to avoid wasting resources on account of attempting to develop the maintainer genotype from plants with Owen cytoplasm.     

phaG和phaC基因在烟草叶绿体中的转化及其遗传分析

中长链羟基脂肪酸聚酯(medium-chain-length-PHAs, mcl-PHAs) 属于微生物聚酯。羟酰-CoA-ACP-转移酶和II型PHA合酶是mcl-PHAs生物合成途径中的两个关键酶。将编码羟酰-CoA-ACP-转移酶的基因phaG与水稻叶绿体psbA基因的启动子和终止子连接构建表达盒RpsbA-pro-phaG-RpsbA-ter,将II型PHA合酶的基因phaC与水稻叶绿体16S rRNA基因的强启动子Prrn及rbcL基因的终止子连接构建表达盒prrn-phaC-RrbcL-ter,连同壮观霉素抗性基因aadA表达盒prrn-aadA-TpsbA-ter一起克隆进烟草叶绿体基因组同源片段rbcL和accD之间,得到烟草叶绿体表达载体pTGC。用包裹有质粒pTGC的金粉子弹轰击烟草无菌苗叶片,经壮观霉素筛选后获得6株叶绿体型转基因植株。对T₀代和T₁代转基因植株进行PCR检测和Southern blot分析表明,外源基因已整合进烟草叶绿体基因组中,且T₁代转基因植株已同质化。RT-PCR分析结果证实外源基因已在转录水平上表达。转基因植株的自交及正反交结果表明,外源基因在转基因后代中能够稳定遗传,遗传方式遵循母性遗传规律,不存在转基因的花粉漂移现象。