中华补血草Syntaxin基因的克隆(英文)

[目的]对中华补血草Syntaxin基因进行克隆。[方法]以中华补血草叶片为材料,提取总RNA并进行反转录反应。依据已知Syntaxin基因5’端EST序列信息设计巢式引物,以反转录得到的cDNA为模板,利用2轮PCR扩增cDNA3’末端(3’RACE),获得Syntaxin基因3’端序列。[结果]获得1090bp的DNA片段。分析表明,该片段编码LsSyntaxin全长基因,其中开放阅读框长816bp,编码271个氨基酸。该基因编码的Syntaxin蛋白相对分子量为30254.3Da,理论等电点为5.55。[结论]为研究Syntaxin基因在补血草中的功能及其在补血草泌盐过程的作用奠定了基础。     

The frost tolerance of Miscanthus at the juvenile stage: Differences between clones are influenced by leaf-stage and acclimation

Miscanthus, a perennial, C₄ grass, has numerous advantages for biomass production, notably its high yield per hectare and low input requirements. However, establishment of this crop may be affected in Europe by frost damage when stem emergence occurs early in the year. The principal aim of this study was to quantify the impact of frost on young miscanthus shoots at different leaf-stages, and to characterise inter-species variations in frost tolerance. Four clones belonging to two species were tested at three leaf stages in a climate-controlled chamber simulating acclimation conditions and frost treatments. Frost tolerance was scored using a 0-6 visual assessment scale and analysed with nonparametric tests.We were thus able to show that more developed plants (6 or 7-leaf stage) were less frost tolerant than those at the 3 or 5-leaf stage. Plants at the 6 or 7-leaf stage also displayed differences in tolerance between clones. The leaf-stage of the plant is linked to apex height, and this appeared to play a role in frost tolerance. M. sinensis displayed variable frost tolerance (tolerance score of between 3.6 and 4.9), although the three clones observed were always more frost tolerant than M. x giganteus (with a score of 3). Moreover, the differences in frost tolerance were negatively correlated (r = −0.94) with the mean leaf surface area of clones at the time of frost exposure. Finally, we observed that acclimation at 12 °C under strong light intensity (600 μmol m⁻² s⁻¹) enabled an increase in the tolerance of young shoots in all the clones tested.     

六道木嫩芽快速繁殖的研究

为满足观赏栽培对六道木种苗的需求,以具生长点的嫩茎为材料,采用组织培养的方法,进行了生长点的生长、生长芽的分化、分化芽的生根、试管苗的生根及生根继代增殖培养、试管苗的移栽与定植的研究。结果表明：1／2Ms＋6-BA0．8mg／L＋NAA0．1mg／L是生长点伸长生长培养的理想培养基;MS＋AgNO30．5mg／L＋ZT1．5mg／L＋NAA0．1mg／1．是生长芽分化培养与分化继代增殖培养的理想培养基;1／4MS＋蔗糖15g／L＋ABT0．9mg／L是分化芽生根培养和试管苗生根继代增殖培养的理想培养基。试管苗移栽成活率为95．6％,定植成活率为98．4％,定植的试管苗保持了六道木的植物学性状和观赏性状。     

苋菜MDH基因克隆及生物信息学分析

[目的]对苋菜MDH基因进行克隆及生物信息学分析.[方法]以苋菜试管苗为材料,从实验室构建的苋菜转录组数据库中筛选出苋菜MDH基因的cDNA序列片段,采用RT-PCR技术对其ORF进行克隆,并进行生物信息学分析.[结果]MDH基因编码344个氨基酸,通过生物信息学分析表明,MDH可能为非分泌蛋白,包括33个磷酸化位点,...     

香蕉果实类受体蛋白激酶基因克隆及RNAi载体构建

植物类受体蛋白激酶在植物的生长发育和信号转导中起到重要作用。以香蕉果实cDNA噬菌体文库为材料,通过原位杂交方法获得1个长度为1 689 bp的香蕉果实类受体蛋白激酶基因,编码563个氨基酸序列,包含1个高度保守的蛋白激酶家族结构域。Southern杂交分析结果表明,该基因在香蕉基因组中不止有1个拷贝。构建了该基因的RNAi干扰载体,为该基因功能研究打下基础。     

新疆三芒草DREB基因片段的克隆

[目的]克隆新疆三芒草DREB基因片段。[方法]利用PCR技术和RACE技术,从新疆沙漠边缘抗旱植物三芒草中克隆抗旱相关基因DREB片段。[结果]研究克隆到了4个DREB基因片段。[结论]该研究为进一步获得DREB基因全序列和研究植物抗旱分子机理、培养抗旱作物品种奠定了基础。     

黑木相思17号无性系在桂东地区早期生长表现

为丰富桂东地区速生树种多样性以及为生态敏感地区桉树改造提供新的树种,2017年5月在桂东地区试种黑木相思17号无性系20hm^2,分别在造林后第8个月和14个月调查试验林不同海拔、坡向、坡位的林分生长情况,试验结果表明:黑木相思17号无性系在桂东地区造林成活率85%以上,造林后第8个月成活率、地径和树高分别为92%,3.1cm和2.3m,造林第14个月地径、胸径和树高分别为7.4cm,5.8cm和5.0m;黑木相思造林后8-14个月,上坡生长表现优于下坡,海拔200m生长表现优于海拔320m。说明黑木相思17号无性系适合在桂东地区生长,避开谷底和山脚等光照不足地块、及时抚育是黑木相思在桂东地区造林成功的关键。     

Clone distribution of the earthworm Eiseniella tetraedra (Sav.) (Oligochaeta: Lumbricidae) across an altitudinal gradient on subarctic mountains of NW Europe

In earlier studies, we have shown that clone diversity of the parthenogenetic earthworm Eiseniella tetraedra increases from the upper reaches of rivers in northern Sweden towards their mouths. Now we survey brooks in the Scandes Mountains in the watershed between Sweden and Norway where major rivers originate. Using starch gel enzyme electrophoresis, we found 37 clones in a total catch of 379 individuals from six mountains. The most abundant clone made up 48.3% of the individuals collected. It was present on most mountains and was found at different elevations. In comparison with other clones it may represent a general-purpose genotype adapted to environmental conditions ranging from alpine through to subalpine to boreal habitats in the mountains. Diversity of clone assemblages decreased with increasing elevation. On four mountains, one to two clones were found at higher elevations. Passive downstream dispersal of E. tetraedra propagules from wider areas of the mountains was responsible for the more diverse clone pools in the lower reaches of the brooks (i.e., “small rivers behave like large ones”). Two clone groups, which deviated from the norm clone in their number of enzyme variants, were evenly distributed among different elevations. Therefore, we could not correlate genotype differences (i.e., adaptation of clones to mountain elevations). Clone pool similarities among the mountains were low on average (range 0–58%) but in a cluster of four mountains, similarities varied from 46% to 58%. Clone pool similarities between different elevations of the same mountain ranged from 27% to 83%. One mountain brook was sampled over 3 years to assess clone turnover. Only the norm clone was found in upstream habitats but it and three other clones were recorded downstream in at least 2 years. Ten clones were found once in the latter habitat as well.     

小麦TaCYP78A5基因的克隆及生物信息学分析

细胞色素P450(Cytochromes P450)蛋白家族含有硫羟基和血红素结构域,参与多种生物合成,CYP78A家族成员对籽粒大小形成有重要的功能。利用同源克隆的方法,分别从小麦大粒品系‘P271’和小粒材料‘中国春’中扩增到位于2AS、2BS、2DS上的TaCYP78A5-2A、TaCYP78A5-2B和TaCYP78A5-2D基因,编码534、544、549个氨基酸。通过分析发现在‘P271’和‘中国春’克隆到的TaCYP78A5序列一致,说明TaCYP78A5基因在这2个粒型的小麦中是保守的。CYP78A家族在水稻和拟南芥中具有非自主性细胞表达模式,结合对CYP78A家族的生物进化关系研究发现,CYP78A家族的各个基因在单子叶植物中是保守的。‘P271’和‘中国春’籽粒大小的差异,可能是TaCYP78A5基因在‘P271’中的基因表达量高于‘中国春’,引起种子发育过程中内表皮细胞的增殖,从而促进种子表皮增大,使得‘P271’种子大小和质量增加。     

贝莱斯芽孢杆菌的内切葡聚糖酶基因的克隆、表达及其酶学性质分析

本实验利用PCR方法扩增1株贝莱斯芽孢杆菌(Bacillus velezensis)源的内切葡聚糖酶基因(CMCase),插入到p ET32a(+)中构建重组表达大肠杆菌系统,对重组内切葡聚糖酶基因进行生物信息学分析以及酶学性质研究。结果表明:内切葡聚糖酶由499个氨基酸组成,预测分子量为55.02 ku,最大开放阅读框ORF约为1 500 bp。经诱导表达优化后,培养上清中可检测到内切葡聚糖酶酶活力为5.81 IU/mL,菌体中为1.61 IU/mL,诱导后原菌液直接超声破碎处理可达7.41 IU/mL,其酶活力为野生菌株的2.3倍。重组内切葡聚糖酶具有典型内切纤维素酶的特征,其最适酶反应温度和pH分别为50℃和6,在60℃条件下放置1 h相对剩余酶活力可保持在70%以上,在pH 6～10范围内可保持90%以上。Na~+、Mg~(2+)可以促进重组内切葡聚糖酶酶活力,而Co~(2+)、Hg~(2+)、Fe~(2+)等金属离子以及表面活性剂SDS则具有较强的抑制作用。由此可见,本实验在大肠杆菌BL21(DE3)中成功表达了内切葡聚糖酶,且该酶主要进行分泌表达,具有一定的耐热性和良好的pH稳定性。