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[目的]了解马尾松干旱胁迫转录组序列的功能分布和SSR位点分布特征,并探索与干旱关联的SSR位点。[方法]对马尾松幼苗进行持续干旱胁迫,选取干旱10、15、25 d及正常供水对照的马尾松针叶样品,通过总RNA提取、Illumina测序、raw reads去冗、Trinity拼接获得unigenes。利用Blast比对,对Unigene序列进行GO、KOG、KEGG功能注释及分类;利用Misa软件进行SSR位点批量搜寻,primer 3. 0软件进行规模化SSR引物设计;利用GOSeq(1. 10. 0)、KOBAS(v2. 0. 12)软件对差异表达的含SSR位点Unigene进行GO、KEGG显著性富集分析。[结果]马尾松转录组194 821个Unigene中,101 806个Unigene获得注释,包含64 943个GO功能注释、35 880个KOG功能注释及30 882个KEGG注释。搜寻到6 728个SSR位点,分布于6 367个Unigene中,SSR出现频率为3. 45%;重复类型以单、三、二核苷酸为主,分别占总SSR的35. 82%、33. 03%和25. 22%;重复基序以A/T、AT/AT、AG/CT、AGC/CTG、AAG/CTT为主;基序长度以10 20 bp的短序列SSR为主;基序重复次数以5～10次重复占优势;批量设计13 338对SSR引物。422个含SSR位点Unigene具有差异表达; KEGG富集分析发现,有11个含SSR位点的差异Unigene参与了光合作用、植物激素信号传导及类胡萝卜素合成等3个与干旱响应相关的代谢途径。[结论]从较高质量的马尾松转录组中获得101 806个具有注释的Unigene;从6 367个Unigene中挖掘出6 728个SSR位点; 422个含SSR位点的差异表达Unigene中筛选出11个与干旱关联的SSR功能位点,为抗旱功能基因定位及马尾松抗旱分子机制研究奠定基础。 相似文献
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[目的]全面了解滇东南特有濒危植物长梗杜鹃转录组SSR位点的分布及序列特征,为长梗杜鹃的保护和合理开发利用提供遗传学资料,为同属植物及近缘种SSR标记的开发及遗传研究提供便利。[方法]利用Illumina Hiseq 4000高通量测序平台对长梗杜鹃叶片进行转录组测序,再通过MISA软件对测序所得Unigenes进行SSR位点的发掘和分析。[结果]发现含SSR的序列17 354条,共得到23 192个SSR,出现频率为31.30%,平均每3 kb出现1个SSR。二碱基和三碱基重复为长梗杜鹃SSR主要重复单元类型,分别占SSR总数的69.25%和15.07%,187种重复基元中,所占比例最高的是(AG/CT)n(62.01%),其次是(A/T)n(12.34%)、(AC/GT)n(4.52%)和(AAG/CTT)n(4.23%)。在SSR和CDS的交集基因中,共发现15 908个SSR位点,其中2 792个位于编码区,出现频率为0.076 SSR/kb,而非编码区为0.344 SSR/kb,在基因编码区中出现频率最高的是三碱基重复(1 356,48.57%)。在不同长度重复单元中,二碱基重复SSR长度变异程度最高,其次是单碱基重复。长梗杜鹃SSR的频率和长度呈显著负相关(P0.01),相关系数为-0.566。[结论]长梗杜鹃转录组SSR位点的出现频率高、分布密度大、基元类型丰富、重复次数较高、长片段较多,具有较高的多态性潜能,用于遗传分析的潜力很大,能满足该物种的保护遗传学研究。 相似文献
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A normal variety of rice (Oryza sativa L.cv. Taichung 65, T65c), its isogenic dwarf line (T65d
1), and a semidwarf variety of a different line (Oryza sativa L.cv. IR8, IR8) were studied. The results were compared with those of an isogenic dwarf line (Rh
i) of wheat straw, which was previously reported. Expression of the dwarf gene,d
1, on the chemical composition and the structural features of lignin present in rice internodes differs from that in an isogenic dwarf line of wheat. The differences include the lignin content, total yield of alkaline nitrobenzene oxidation products, and distribution of wall-bound hydroxycinnamic acids. There was, however, no difference in the syringyl/ guaiacyl nuclei (S/V) molar ratio and neutral sugar composition. The lignin composition of rice straw cell walls, particularly that of the dwarf variety, contained more of the condensed structure and fewer syringyl nuclei than lignin in wheat straw cell walls. It is suggested that crosslinking between lignin and polysaccharides by ester-ether bridges via ferulic acid contributes to the mechanical properties of the cell walls of rice straw. Thus the chemical and structural characteristics of lignin in rice straw differ to some extent from those of other temperate grasses, such as wheat (Triticum aestivum) and phalaris (Phalaris aquatica), as reported previously. This can probably be attributed to the water environment of rapidly growing rice seedlings, but it also depends on the genetic variety of the rice plant. 相似文献
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A transformation system was established for loblolly pine (Pinus taeda L.) mature zygotic embryos usingAgrobacterium tumefaciens. The gene coding for the β-glucuronidase (GUS) gene was introduced into loblolly pine tissues and its transient expression
was detected with histochemical staining. The influences of different genotypes.Agrobacterum concentrations. and cocultivation time on GUS expression and kanamycin resistant callus and shoot regeneration were investigated.
The results showed that the highest GUS expression frequency (16.3%) and shoot regeneration frequency (78%) were obtained
from genotype 9–1003 withAgrobacterium concentration decreased 9 times and cocultivation time of 56 hours respectively. GUS expression was obtained in all genotypes
tested. The successful expression of the GUS gene in different genotypes suggested that it will be a useful transformation
system for loblolly pine.
(Responsible Editor: Chai Ruihai) 相似文献
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生长素及其信号转导系统对植物的生长发育具有重要的影响。本研究从银腺杨'84K'(Populus alba × P. glandulosa cl. '84K')中分离了生长素受体基因PtrFBL1,利用PMDC32构建了PMDC32-PtrFBL1超量表达载体,并通过遗传转化获得了超量表达植株17个。对温室定植的3个转基因株系和对照植株的根系、生长量和光合指标等性状分析结果显示:转基因株系总根长和总根面积达到显著或极显著差异,而根系干质量、平均不定根系长度、平均不定根直径差异不显著;株高、平均节间长、地径和高径比皆高于对照,且大多数转基因株系达到显著差异;除气孔限制值(Ls)低于对照外,气孔导度(Cd)、水分利用效率(WUE)、光能利用效率(LUE)和叶绿素相对含量皆高于对照,且大多数转基因株系达到显著或极显著差异。以上结果表明,可能是FBL1超表达增加了转基因株系根系面积,提高了水分和养分的吸收利用,进而导致转基因株系光能吸收和转化效率提高,引起转基因株系生长加快。 相似文献
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【目的】为研究杧果树体发育过程中GA合成调控以及GA信号转导的作用提供参考,并为矮生型杧果品种及矮化砧木的选育提供参考。【方法】以广西地方具有矮化特性杧果品种‘桂热杧82号’无病虫侵害的嫩叶为材料,对其GA2ox基因克隆,并进行亚细胞定位及表达分析。通过改良Trizol法提取叶片总RNA,利用简并引物克隆GA2ox基因片段,采用RACE法得到基因的全长cDNA序列,命名为MiGA2ox。通过生物信息学方法分析其结构特征,利用GFP进行亚细胞定位,采用实时荧光定量RT-PCR技术研究MiGA2ox基因在‘桂热杧82号’与乔化品种‘金煌杧’中的表达特性。【结果】MiGA2ox全长cDNA序列为2 051 bp,含有3′非翻译区约900 bp,5′非翻译区约400 bp。GA2ox序列均含有完整的开放阅读框,终止密码子为TGA,编码341个氨基酸,与橙子(XM_006467404.3)、柑橘(XM_006449629.2)、木薯(XM_021738345.1)、巴西橡胶树(XM_021820571.1)、陆地棉(XM_016896568.1)、中棉(XM_012597405.1)的一致性分别为79%、79%、78%、78%、76%、75%。MiGA2ox编码蛋白的相对分子质量为38 729.4 Da,等电点为6.56,为稳定蛋白,无信号肽,无明显疏水区,无跨膜结构域。根据共聚焦激光扫描显微镜观察结果可知,MiGA2ox编码蛋白定位于细胞核中。在随机选择2个时间节点,矮化品种‘桂热杧82号’MiGA2ox的表达量均高于乔化品种‘金煌杧’。【结论】‘桂热杧82号’MiGA2ox基因的表达与其矮化性状有关。 相似文献
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Katsuaki Ishii 《Journal of Forest Research》2002,7(2):99-104
Hinoki cypress (Chamaecyparis obtusa) is one of the most important timber resource forest trees in Japan. Because seed production from a seed orchard of hinoki
cypress is not constant every year, micropropagation from a limited amount of material is useful. Up to now, the conventional
tissue culture method using solid medium has been used. Here a new method using liquid culture in tubes rotated vertically
is described. Shoot primordium of hinoki cypress was inoculated in Campbell and Durzan’s (CD) liquid medium containing different
cytokinins (6-benzylaminopurine (BAP), Zeatin, thidiazurone (TDZ)), and the container tubes were rotated vertically around
the axis at 2 times / min. Culture room temperature was 25°C and light condition was 16 h photoperiod per day of fluorescent
lamps. Zeatin at 1μM concentration was the best for maintaining the shoot primordium production and TDZ induced callus on the surface of the
shoot primordia. After shoot primordium multiplication in the liquid culture, they were transplanted to agar medium for shoot
elongation. A high concentration of agar (up to 16 g/L) or AVF (anti vitrification factor from Dr. Nairn, 1995) was effective
to prevent vitrification of the shoots. Transformation of shoot primordium was done using particle bombardment with vectors
containingβ-glucuronidase (GUS) gene or herbicide resistance gene (bar). Positive result for transient transformation was observed with the histo-chemical study for transformation with GUS. Integration
of a useful herbicidebar gene into the shoot primordium culture system was also tried and stably transformed plants were obtained. This is the first
report of stable transformation of Japanese conifer using practically useful gene.
The generous supply of AVF-B from Dr. B.J. Nairn, Tasman Forestry, NZ is also appreciated. 相似文献
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拟南芥 AtFBDL1 基因是FBD-like基因家族的一员,其编码蛋白含有类似于F-box的结构域。表达模式网络预测结果显示该基因在茎顶端分生组织中高丰度表达,但对于FBD-like基因家族的研究还很少,其功能目前尚不明确。为此,本研究通过组织半定量表达分析和GUS染色显示 AtFBDL1 基因在拟南芥中具有时空表达特异性。结果表明:在真叶形成前和形成初期,该基因主要在茎顶端分生组织和下胚轴区域表达;真叶形成后,该基因在下胚轴的表达明显减少,而主要集中在茎顶端分生组织表达。遗传转化显示:与野生型植株相比,过表达 AtFBDL1 基因的植株生长发育缓慢,抽薹时间推迟3 4 d,莲座叶叶片面积减小,叶片数目平均增多10片,并且伴随有变态叶出现;过表达植株株高比野生型矮,株高最大差值达到12 cm。共表达网络预测 AtFBDL1 与多个与生长素和花发育相关的基因具有共表达关系。以上研究结果表明: AtFBDL1 基因在拟南芥的生长发育过程中,特别是在顶端分生组织分化过程中起重要作用。 相似文献
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油桐尺蛾核型多角体病毒广西株基因组序列由Sanger测序的方法得到。拼接后获得了121 268 bp序列,GC含量为36.76%。以长度不小于50 aa的序列认定为功能基因,共预测到131个开放阅读框,通过Blast比对,其中87个能注释到功能基因。对基因组序列进重复序列分析,共发现SSR位点174个,同源重复序列1个。在SSR位点中,包含6种长度的重复基序,且以富含AT的重复基序为主。同源重复序列两端为一59 bp片段或其一部分组成的重复序列,两端分别重复12次和7次,中间则为一段与重复片段无关的236 bp片段。与武汉株相比,广西株病毒在37个核心基因中有4个存在蛋白序列长度上的差异,而在其他非核心基因中有6个基因仅存在于一个病毒株中,另有13个基因存在蛋白序列长度上的差异。 相似文献