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.     

菊苣叶绿体同源片段的克隆及多顺反子定点整合表达载体的构建

利用叶绿体基因组进化中高度保守的特点,根据烟草叶绿体基因组全序列设计合成引物,从菊苣叶绿体基因组中扩增包括rps7-rps12-trnV-16S rDNA基因在内的一段序列(GenBank登录号为GQ199478),并以此作为定点整合外源基因的同源重组片段。以叶绿体基因强启动子prrn驱动选择标记基因aadA及报告基因gfp构建多顺反子表达盒prrn-aadA-gfp-psbA-3,′然后将表达盒克隆进菊苣叶绿体同源片段中,获得菊苣叶绿体定点整合表达载体pJAG,酶切分析证明所构建的载体符合预期设计。该载体的构建为建立菊苣的叶绿体转化体系奠定基础,为进一步通过叶绿体基因工程手段将更多感兴趣的基因导入菊苣进行遗传改良,或以菊苣叶绿体作生物反应器生产动物口服疫苗等搭建技术平台。     

蓝光对水稻幼苗叶绿体发育的影响

蓝光对水稻幼苗叶绿体发育的影响李韶山，潘瑞炽（华南师范大学广州５１０６３１）关键词：蓝光，叶绿体，黄化质体，转绿，类囊体ＥｆｆｅｃｔｏｆＢｌｕｅＬｉｇｈｔｏｎｔｈｅＤｅｖｅｌｏｐｍｅｎｔｏｆＣｈｌｏｒｏｐｌａｓｔｓｉｎＲｉｃｅＳｅｅｄｌｉｎｇｓ￥ＬＩ...     

油菜叶绿体同工酶的研究

本实验采用聚丙烯酰胺凝胶盘状电泳的方法和酶电游图谱扫描以及对酶液中酶活和蛋白质含量的测定,研究了杂种油菜F_1(S45×81008—34)及其亲本在初花期的叶绿体过氧化物酶同工酶。结果表明:(1)过氧化物酶同工酶在叶绿体中是真实存在的。(2)F_1叶绿体的过氧化物酶同工酶图谱显示出杂种油菜F_1不仅具有它的亲本的互补酶而且还具有它的亲本没有的杂种酶,它明显地表现出杂种优势。(3)杂种油菜F_1的酶液中的酶活及蛋白质的含量皆高于它的亲本的酶活和蛋白质含量。     

植物生长调节剂“784—1”对大豆叶肉细胞超微结构的影响

本文应用植物生长调节剂“784—1”进行大豆种子浸种和拌种处理。田间试验与电镜观察结果表明,与对照组比较,叶肉细胞中的线粒体、叶绿体和叶绿体内片层结构呈增多趋势。     

Chloroplast DNA variation in European potato cultivars

Summary The chloroplast DNA of 27 potato cultivars and 2 wild species has been examined by Southern blotting using probes representing the entire chloroplast genome of barley. 16% of the probe × enzyme combinations detected polymorphism between the potato cultivars. Cvs Maris Piper, Croft, Shelagh, Stormont Enterprise and Estima and the wild species could be distinguished from the other cultivars by their chloroplast DNA restriction patterns. These results are discussed in relation to cytoplasmic diversity and potato improvement.     

高粱5-烯醇式丙酮酰莽草酸-3-磷酸合酶基因(EPSPS)叶绿体转运肽(CTP)的克隆及其在转基因玉米中的功能验证

5-烯醇式丙酮酰莽草酸-3-磷酸合酶(EPSPS)催化莽草酸-3-磷酸(S3P)与磷酸烯醇式丙酮酸(PEP)合成5-烯醇式丙酮酰莽草酸-3-磷酸(EPSP),具有与草甘膦结合的活性位点,且结合后会抑制EPSPS的活性,在植物抗除草剂基因工程中具有重要的应用价值.为了培育抗草甘膦玉米(Zea mays),本研究通过对高粱(Sorghum bicolor)EPSPS基因的结构分析,克隆了该基因5'端的叶绿体转运肽(chloroplast transit peptide,CTP).将该转运肽与来源于农杆菌(Agrobacterium tumefaciens)菌株CP4 EPSPS基因整合,以Ubiquitin为启动子,35S polyA为终止子,构建表达载体,同时以不含有转运肽的CP4 EPSPS基因为对照,遗传转化玉米得到稳定转基因株系;用PCR、Southern blot、ELISA等方法检测转基因玉米CP4 EPSPS基因的表达量并对其进行草甘膦抗性检测,研究发现,不含转运肽的转化事件虽然CP4 EPSPS基因表达量与含有转运肽的基本一致但却不具有草甘膦抗性,而含有转运肽的转化事件则抗性明显.说明转运肽并不影响CP4 EPSPS基因的表达,但对转基因植株草甘膦抗性起着重要作用,说明本研究克隆的叶绿体转运肽能够正常行使其生物学功能.研究结果为利用EPSPS基因培育抗草甘膦作物提供了重要参考资料.     

Maternally and paternally inherited molecular markers elucidate population patterns and inferred dispersal processes on a small scale within a subalpine stand of Norway spruce (Picea abies [L.] Karst.)

The within-population spatial structure of genetic diversity is shaped by demographic processes, including historical accidents such as forest perturbations. Information drawn from the analysis of the spatial distribution of genetic diversity is therefore inherently linked to demographic-historical processes that ultimately determine the fate of populations.All adult trees and saplings in a 1.4-ha plot within a mixed Norway spruce (Picea abies [L.] Karst) stand were characterised by means of chloroplast (paternally inherited) markers, and a large sub-sample of these were genotyped at mitochondrial (maternally inherited) molecular markers. These data were used to analyse the spatial distribution of genetic variation and to compare the patterns corresponding to the two marker types. The plot presented non-homogeneous local stem density in the younger cohorts, and the indirect effect of this source of variation on the spatial genetic structure was investigated. Results suggest that (i) spatially limited seed dispersal induced patchiness in genotype distribution, while pollen flow had a homogenising effect; (ii) deviations from random spatial structure were stronger in the demographically most stable portions of the stand, while they were weaker where sudden bursts of regeneration occurred; (iii) spatially overlapping adult and sapling cohorts displayed the same spatial genetic structure (stronger on stable areas, weaker in portions of the stand undergoing events of intense regeneration), which was substantiated by the influence of local demographic processes. Regeneration dynamics as modulated by demography thus influences the distribution of genetic diversity within the stand both in the younger life stages and in the adult population.     

水稻巯基蛋白酶抑制剂基因在烟草叶绿体中的表达

将水稻巯基蛋白酶抑制剂(Oryzacystatin,OC) cDNA基因克隆、测序分析后,与烟草(Nicotiana tabacum L.)叶绿体16S rDNA基因启动子和T393终止子构建表达盒,与抗壮观霉素选择标记基因aadA表达盒相连,以烟草叶绿体基因组同源片段rbcL和ORF512一起构建成叶绿体转化载体pZOC.通过基因枪轰击烟草幼苗叶片,壮观霉素筛选,获得转化再生