等渗Ca和NaCl胁迫对黄瓜叶片叶绿体超微结构及光合特性的影响

采用营养液栽培,以盐敏感型的黄瓜品种津春2号为试材,研究了等渗Ca和NaCl胁迫对黄瓜幼苗叶绿体超微结构、叶片叶绿素含量、光合特性和叶绿素荧光参数的影响。结果表明：两种盐胁迫均降低了黄瓜叶片叶绿素含量,且NaCl胁迫后下降的幅度大于Ca胁迫。Ca胁迫后,叶绿体大部分与细胞壁脱离,基粒片层数减少|等渗的NaCl胁迫后,叶绿体结构变形,基粒片层膨胀、松散,片层之间界限模糊,嗜饿颗粒明显增多。Ca胁迫后净光合速率（Pn）、气孔导度（Gs）、PSⅡ最大潜在光化学效率（Fv/Fm）、PSⅡ实际光化学效率（ΦPSⅡ）和光化学淬灭系数（qP）下降的幅度均小于NaCl胁迫。Ca胁迫下植株的胞间CO2浓度（Ci）呈下降趋势,而NaCl胁迫下Ci呈上升趋势。综上所述,Ca胁迫主要通过气孔限制引起Pn下降|而NaCl胁迫严重破坏了叶绿体超微结构,主要通过非气孔因素限制引起Pn下降。     

叶绿体型转昆虫抗冻蛋白基因烟草的耐寒性

根据已构建的大豆叶绿体表达载体pJY01,设计特异性引物,将昆虫抗冻蛋白基因MpAFP149插入此载体中构成叶绿体表达载体pJY01-MpAFP149,利用基因枪轰击法转化烟草,经壮观霉素筛选获得4株叶绿体型转抗冻蛋白基因烟草株系。PCR和PCR-Southern结果显示外源基因已整合至烟草叶绿体基因组中但同质化水平不高,RT-PCR结果也表明昆虫抗冻蛋白基因已发生了转录。将野生型烟草、叶绿体型转抗冻蛋白基因烟草及核转化T₁代转抗冻蛋白基因烟草(pCAMBIA1302- MpAFP149)于–1℃低温处理3 d,观察耐寒表型及测定相对电导率。结果表明, 叶绿体型转基因烟草的耐寒表型优于野生型烟草,但与核转化的T₁代转抗冻蛋白基因烟草无显著差异。处理3 d时,叶绿体型转基因烟草和T₁代转抗冻蛋白基因烟草的电导率分别为39.2%和38.2%,而野生型烟草已达73.7%。本实验获得的异质化转叶绿体抗冻蛋白基因烟草与转核基因烟草的耐寒力无差异。     

二穗短柄草叶绿体RNA编辑位点的预测及其鉴定

RNA编辑是陆生植物叶绿体转录后基因表达调控的一种重要方式。为进一步探讨单子叶植物RNA编辑功能及其发生机制,本研究通过生物信息学方法,对二穗短柄草叶绿体的81个蛋白编码基因的RNA编辑位点进行了预测和鉴定分析,结果发现78个基因存在编辑位点,共检测到176个编辑位点,都是C到U的转换。其中ndhB最多,为14个编辑位点。为验证预测结果,利用blast工具比对了NCBI的二穗短柄草EST数据库,最终确定存在于17个基因中的34个编辑位点是真实存在的,其中19个为沉默编辑,15个为有效编辑。与5个不同单子叶禾本科物种18个蛋白编码基因的RNA编辑位点的比较发现,在rpoB-206位点,只有二穗短柄草发生了编辑,且只有ndhD-295(293)为部分编辑位点。     

蝴蝶兰叶绿体DNA微卫星分析与标记开发

为开发兰科植物叶绿体微卫星(cpSSR)引物,对台湾蝴蝶兰叶绿体全序列进行cpSSR统计分析。结果表明台湾蝴蝶兰SSR以单碱基重复为主,占总数的78.3%。在单碱基重复中又以A和T重复为主,占95.8%。重复方式以完全重复为主,占总数的70%。开发出3对cpSSR引物。     

枇杷叶片气孔保卫细胞叶绿体数目分析

[目的]研究枇杷叶片保卫细胞的叶绿体数目差异,探讨其变化规律。[方法]取43份枇杷资源嫩叶进行处理,在20倍显微镜下进行统计,随机选择50个保卫细胞计算叶绿体数目。[结果]枇杷叶片保卫细胞叶绿体数目在11～17,平均为13.19个,品种间差异大。不同品种叶绿体数目变异幅度不同,在9～14,仅栎叶枇杷的叶绿体数目大于16。[结论]栎叶枇杷可能是多倍体,这可为枇杷资源的倍性鉴定提供参考。     

小麦叶绿素缺失突变体Mt135的叶绿体基因差异表达分析

小麦叶绿素缺失突变体Mt135自交后代稳定表现绿株、条纹株和白化株3种类型, 其中条纹株白色组织和白化株的叶绿体数目和结构发生突变, 完全失去光合能力。为研究该突变体叶绿体基因表达与光合作用的关系, 采用实时荧光定量PCR技术, 分析了白化株和条纹株的叶绿体基因表达。在白化株中共检测到40个差异表达基因, 涉及4类功能(编码光反应相关蛋白、编码叶绿体内能量代谢相关酶、核糖体合成和tRNA合成), 包括18个上调表达和22个下调表达基因;在条纹株中共检测到13个上调表达基因, 其表达变化趋势与在白化株中一致。白化株的差异表达基因中, 编码光系统II、I结构蛋白的psb、psa及ycf等基因家族的基因表达量显著下调;多个编码核糖体蛋白大、小亚基的基因表达量改变, 尤其是核糖体蛋白小亚基编码基因rps14和23S rRNA的编码基因23S rDNA表达量显著下调。推测Mt135突变性状与参与光反应相关蛋白的编码基因、叶绿体内能量代谢相关酶的编码基因、核糖体合成相关基因以及tRNA合成相关基因表达量的改变密切相关。     

部分箬竹属植物的叶绿体DNA分析（摘要）

[目的]研究箬竹属植物和赤族属植物的亲缘关系。[方法]以箬竹属13个竹种及其近缘的赤竹属3个竹种为材料,采用PCR方法扩增叶绿体trnL-trnF间隔区基因片段,并对其进行序列分析,构建系统树。[结果]利用已发表的trnL-trnF序列通用引物扩增出长度为1008～1103bp的trnL-trnF片段,比较长度为940bp。cpDNA序列聚类将箬竹属与赤竹属竹种混合聚在一起,同源性为99%以上,可分为五组。其中,髯毛箬竹、广东箬竹、小叶箬竹、华箬竹、毛鞘箬竹、矮箬竹、胜利箬竹、箬竹、箬叶竹、粽巴箬竹、翠竹和菲白竹12个竹种聚为一组;泡箬竹、天目箬竹、阔叶箬竹和美丽箬竹4个竹种各自成一组。[结论]竹种间的同源性极高表现为较慢的进化速率,提供的信息位点很少,不能很好地解决箬竹属种间的系统学问题。     

水稻条斑花叶突变体生态st(t) 的鉴定与遗传定位

利用EMS诱变育成优良籼型恢复系缙恢10号,从其后代中鉴定出一个白色条斑花叶突变体st(t),在三叶期开始表现白斑,拔节期白斑变为不规则线状,一直保持到成熟。突变体叶绿素含量明显下降,类胡萝卜素含量显著升高。透射电镜观察表明,突变体的绿色叶片部位与野生型相比,在细胞结构上无明显差异,叶绿体发育正常;突变体的白化部位细胞结构异常,质体内多含有积聚在一起的嗜锇小球,不能发育出正常叶绿体所具有的类囊体和基质片层结构。遗传分析表明该性状受一对隐性核基因调控,利用1 500株西农1A/st(t)的F₂隐性定位群体,最终把St(t)基因定位在第6染色体SSR标记RM19745和RM19762之间,遗传距离分别为0.07 cM和0.27 cM,根据9311基因组序列推测,两标记之间的物理距离约为345 kb。这为St(t)基因的图位克隆和分子标记辅助育种奠定了基础。     

Molecular polymorphism of cytoplasmic DNA in Ficus carica L.: Insights from non-coding regions of chloroplast DNA

The trnL (UAA) intron and the intergenic spacer between the 3′ exon of trnL (UAA) and trnF (GAA) sequences were used as genetic markers for differentiating Ficus carica cultivars and establishing refined genetic relationships. The study was based on 20 fig cultivars, collected from south and centre of Tunisia. Since, the intron was thought to be more variable among close relatives than is the chloroplast spacer. The size of these non-coding regions varied from 554 to 589 and from 989 to 1022 bases pairs for the intron and the combined sequences correspondingly. The average of GC content was 33.9% and 34.6% in the intron and the combined intron and spacer respectively. High values of A + T contents were detected in both data sets and may explain the high proportions of transversions founded. The observed variation pattern of plastid DNA provides evidence of an important genetic diversity. The overall transition/transversion bias (R) was 0.202 in the intron and 0.27 in the combined regions. The RI index of 0.592 indicates that these combined sequences have clearly more homoplasy then the intron (RI = 0.705) and spacer (RI = 0.777) sequences separately. Phylogenetic trees were generated based on maximum parsimony (MP) and neighbor-joining (NJ) analysis of the chloroplast sequences data. Results proved that a typically continuous genetic diversity characterizes the local fig germplasm. In fact, relationships inferred from the cpDNA analysis suggest several clades, which do not show geographical or tree sex correspondence. Although the level of apparent diversity is considerable, we may conclude that non-coding regions of chloroplast genome provide a new and practical opportunity to evaluate genetic diversity and to discriminate fig cultivars. Revealed cytoplasmic DNA markers are reliable to elaborate a molecular data base to conduct management and breeding programs on local fig germplasm.     

Tetraploid muskmelon alters morphological characteristics and improves fruit quality

Tetraploid muskmelon plants were induced successfully from diploid Cucumis melo inbred M01-3 (2n = 24) by colchicine. The morphological characteristics and fruit qualities of diploid and tetraploid muskmelons were investigated. The results showed that the leaves and flowers of the tetraploid plants were markedly larger, the plants were obviously higher, and the stems were thicker than those of the diploid plants. Transmission electron microscope observation revealed that the numbers of chloroplast, granule and grana, and the length of chloroplast and granule of the tetraploid plant leaves were significantly more or longer than those of the diploid plants. The soluble solid, soluble sugar and vitamin C contents in the tetraploid fruit were distinctly higher than those in the diploid fruit. The weight of the tetraploid fruit was 30% heavier than that of the diploid fruit. And the seeds from the tetraploid plants were broader and thicker than those from the diploid plants. Together, our results suggest that the tetraploid muskmelon exhibited better agronomical characteristics than the diploid muskmelon and the tetraploid muskmelon could be used as the improved variety and a potential germplasm for the development of triploid fruit.