陆地棉胞质雄性不育系与保持系线粒体基因组RFLP分析

【目的】研究棉花细胞质雄性不育（CMS）系与保持系线粒体基因组的差异,为克隆棉花CMS基因奠定基础。【方法】以atpA, atp6, atp9, ccmB, ccmC, ccmFN1, cob, coxI, coxII, coxIII, matR, nad1bc, nad2, nad4, nad5, nad6, nad7c, nad9, rpl5, rrn18等20个线粒体基因保守序列为探针,利用Southern blot方法对棉花细胞质雄性不育系、保持系线粒体基因组进行RFLP分析。【结果】发现atpA、atp9、ccmB、nad1bc、nad6、nad7c、rrn18等基因在棉花不育系和保持系间存在明显RFLP多态性,其中atpA的差异最明显。【结论】atpA是棉花CMS系和保持系线粒体基因组间的一个明显差异位点。CMS系比保持系缺失一个rrn18拷贝。     

棉花DELLA蛋白GhGAI4b基因的克隆及功能初步分析

[目的]DELLA蛋白已在不同的物种中被广泛鉴定,并且功能区高度保守,是GA信号通路中的负调控因子,在棉花纤维细胞起始和发育中起重要的作用.研究棉花中DELLA基因的功能.[方法]从棉花中克隆DELLA蛋白的同源基因GhGAI4b,构建35S:GhGAI4b植物过量表达载体,通过农杆菌侵染花序的方法转入野生型拟南芥中,观察转基因植株的表型.[结果]序列分析表明GhGAI4b全长开放阅读框1 617 bp,编码538个氨基酸,具有DELLA蛋白的典型结构域.采用Clustal X将GhGAI4b的氨基酸序列与其他已知的DELLA蛋白氨基酸序列进行相似性比对,发现GhGAI4b氨基酸序列与陆地棉、拟南芥的相似性较高,达到47;以上.转基因株系表现出叶片颜色深绿,叶片缩小且肥厚,叶片细长呈锯齿状;有些株系会出现多重莲座叶,雄蕊发育不全,晚花,株高变矮等赤霉素不敏感突变体的表型.[结论]过表达GhGAI4b能够抑制拟南芥莲座叶的生长和发育,影响转基因拟南芥花器官的发育,严重时引起转基因拟南芥的不育.     

陆地棉单萜合酶基因GhTPS4和GhTPS5的克隆及棉铃虫诱导的基因表达分析

通过本地Blast筛选陆地棉转录组数据并结合RACE的方法,克隆获得2个陆地棉萜类合酶基因全长,命名为GhTPS4(Genebank登录号:KF860865)和GhTPS5(Genebank登录号:KF860866)。GhTPS4和GhTPS5基因分别编码590个和598个氨基酸,预测分子量为68.5 kDa和69.9 kDa,等电点为5.59和5.66。生物信息分析表明:GhTPS4和GhTPS5均含有叶绿体转运肽,并含有"DDxx D"和"RRX8W"保守域,具有典型的单萜合酶特征。此外,聚类分析表明GhTPS4和GhTPS5均属于Tpsb亚家族。利用荧光定量PCR的方法,对GhTPS4和GhTPS5在棉铃虫为害陆地棉(石远321)叶片的表达模式进行了研究,结果表明:GhTPS4的表达量在处理后不同时间均高于未处理对照,在处理后36 h表达量最高,表明GhTPS4在应答棉铃虫为害时可能起到了积极的防御作用。GhTPS5在处理后不同时间均比未处理对照表达量低,其功能尚需进一步探讨。     

Nonstructural carbohydrates in leaves subtending cotton bolls,fibers and embryos in response to nitrogen stress

A field study was conducted in 2013 and 2014 where cotton was exposed to three N regimes: (1) the control without N application (low N); (2) 260 kg N ha^?1 (medium N); (3) 520 kg N ha^?1 (high N). Boll size, lint mass per boll, seed mass per boll, fiber length and strength were significantly decreased under N deprivation in the two years. The increased carbohydrate levels of LSCB (leaf subtending the cotton boll) led to decreased carbohydrate levels of fibers in the low N relative to the other N treatments. The low N embryos exhibited lower starch concentrations at 17 and 31 DPA (days post anthesis), and TNC (total nonstructural carbohydrate) concentrations at 17, 31, 45 and 52 DPA compared to medium N embryos. Starch levels in LSCB had negative associations with those in fibers at 17, 31 and 45 DPA, but positive associations with those in embryos at 24 and 45 DPA. Fibers expressed negative associations with embryos in glucose level at 24 and 38 DPA, and in TNC levels at 17 and 45 DPA. The study suggests that carbon assimilate levels in fibers and embryos could explain the difference in boll yield components and fiber quality.     

棉花黄萎病菌毒素对棉花生化代谢的影响

为明确棉花黄萎病菌毒素对棉花生化代谢的影响,采用棉花黄萎病菌毒素处理棉花黄萎病抗病和感病品种的棉苗后,测定棉株体内超氧化物歧化酶(SOD)、过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)的活性以及丙二醛(MDA)和木质素含量的动态变化。结果表明,抗病品种陕7191对病菌毒素的抗性显著高于感病品种中棉19;接种毒素后2品种棉苗叶片SOD活性都降低,感病品种降幅高于抗病品种;感病品种POD活性升高较快,到达峰值后迅速下降,而抗病品种POD活性持续上升;2品种PAL活性变化趋势相似,接种后抗病品种活性始终高于感病品种;2品种MDA和木质素含量均增多,感病品种MDA含量的增幅和增量高于抗病品种。     

基于异速生长关系的棉籽蛋白质含量模拟模型

基于不同熟性棉花品种的异地分期播种和施氮量试验,运用异速生长模型分析棉籽氮浓度与棉籽生物量间的关系,建立氮素的潜在增长模型,进而综合品种特性、主要气象条件(温度、太阳辐射)和栽培措施(施氮量)等因子,建立基于异速生长关系的棉籽蛋白质含量模拟模型。利用不同生态点品种、播期和施氮量的田间试验资料的检验结果表明:安阳、淮安和徐州试点模拟值与实测值的根均方差(RMSE)分别为2.2%、4.1%和2.3%。说明依据棉籽蛋白质含量与生物量间的异速生长关系模拟棉籽蛋白质形成切实可行,模型预测性好,广适性强。模型的不足在于其精确度受棉籽干物质累积模型和棉铃对位叶N浓度模型的影响较大。     

Genbank��½���ޱ�����б�ǩ(EST)�ͻ��������У�GSS���е�SNP����

陆地棉基因组精细定位需要更加丰富的分子标记,为了阐明单核苷酸多态性(SNP)标记资源开发利用的前景,对GenBank数据库中陆地棉表达序列标签(EST)和基因组序列(GSS)进行分析.下载GenBank数据库中公布的陆地棉EST序列及GSS序列,利用DNAStar软件进行叠连群构建及其候选SNP位点分析.结果表明:陆地棉EST序列307414条及GSS序列242015条,EST序列构建3737个叠连群,序列累计10477241 bp.由4条及以上序列组成的叠连群累计序列长度为3761800 bp,发现候选SNP位点1007258个,叠连群平均出现1个SNP位点最低频率为2.32％.GSS序列共构建1517个叠连群,序列累计1625700 bp,发现SNP位点574296个,叠连群平均出现1个SNP位点最低频率为9.18％.陆地棉的EST和GSS均有频率较高SNP位点,GSS出现SNP频率高于EST序列,开发SNP引物3254对.     

棉花根际解钾细菌的生理活性和促生效果

考察优良棉花根际解钾菌株K1111、K1114、K2115、K3105和K3205的溶磷、解钾、产铁载体和产吲哚乙酸等生理活性。将其接种到棉花根际,通过盆栽试验,研究5株菌株对棉花根系发育与生长的影响。结果表明,这5株菌的产铁载体能力均达到5+(极高量)水平;分泌吲哚乙酸的浓度在6.6～12.4μg/mL之间;同时具有很高的溶磷能力。接种5株菌株的棉花根系活力、株高和干重均显著高于不施钾肥的对照组,高于或相当于施钾对照组。     

Long-Term Effects of Aluminum and Cadmium on Growth,Leaf Anatomy,and Photosynthetic Pigments of Cotton

Aluminum (Al) and cadmium (Cd) are two elements that contaminate soil in different ways as waste products of some industrial processes and that can be tolerated by some plant species in different concentrations. In this study, growth parameters of leaves and stems (fresh and dry weights, stem lengths, leaf surface area, and lamina thickness), anatomical changes in leaves (lower and upper epidermis, stomata and mesophyll tissue), and photosynthetic pigment contents (chlorophyll a and b, total chlorophyll, and carotenoids) were investigated in cotton (Gossypium hirsutum L. cv. Nazilli 84S), which was treated with Al and Cd for 3 months. Cotton seedlings were grown in greenhouse conditions and watered with Hoagland nutrient solutions, which contained 0, 100, and 200 μM aluminum chloride (AlCl₃) and cadmium chloride (CdCl₂). It was observed that reduced soil pH positively affected many parameters in cotton plants. Aluminum accumulation was greater in leaves than stems while the opposite was true for Cd accumulation. Leaves and stems of cotton plants treated with 100 and 200 μM Al and Cd showed slight growth changes; however, high concentrations of Al (200 μM) caused significant reductions in leaf area and leaf fresh weight, whereas stem fresh weight decreased with 200 μM Cd treatment. Anatomical parameters were mostly affected significantly under both concentrations of Al and Cd solutions (100 and 200 μM). The results revealed that the anatomical changes in the leaves varied in both treatments, and the long-term effect of the tested metals did not include harmful effects on anatomical structures. Moreover, the variations could be signals of tolerance or adaptive mechanisms of the leaves under the determined concentrations.     

The effect of starter fertilizers on cotton production in legume residues

Abstract

Field and greenhouse studies were conducted to identify starter fertilizers which would enhance cotton seedling survival, growth, and yield in legume residues. Field studies were initiated in the fall of 1982 on a Norfolk sandy loam (Typic Paleudult) in the Upper Coastal Plain of Alabama. Winter annual legumes, crimson clover (Trifolium incarnatum L.) and hairy vetch (Vicia villosa Roth) were established as whole plots along with a winter fallow area. Split plot treatments consisted of O, N, P, K, NP, NK, and NPK starter fertilizers. The cotton (Gossypium hirsutum L.) was planted with a conservation tillage planting unit with in‐row subsoilers. The starter fertilizers were applied deep (8 to 10 inches) in the subsoil track. Greenhouse studies were also conducted with soil from whole plot areas top dressed with corresponding legume tissue at a rate of .9 g tissue/500 g soil. Seedlings in the greenhouse were rated for disease and emergence, and dry weights were recorded.

Cotton populations in field studies were lower in legume mulched than fallow soils in 1984. Application of starter fertilizers generally increased harvest populations, particularly the NK combination. In 1983, cotton growth was greater in vetch than other soils, but responses to starter fertilizers varied with analyses and years. Seed cotton yields were consistently high with P starter, although P did not always improve cotton stands and growth. When averaged across years and cover crops, yields were 3151, 3031, 2865, 2790, 2753, 2741, 2512, and 2364 for P, NP, P, NP, K, NPK, N and O, starter treatments respectively.

Greenhouse studies indicated that starter fertilizer improved cotton emergence in legume soils, but decreased emergence in fallow soils. Disease ratings of emerged seedlings were more severe when starter fertilizer was used than when it was not used. Thus, starter fertilizer increased emergence and survival, despite high disease ratings. Cotton seedling growth generally increased when poor emergence reduced cotton seedling competition.