转基因棉花抗旱性评价体系的建立以及安全性评价研究

发布时间：2018-03-14 08:32

本文选题：转基因棉花　切入点：干旱胁迫　出处：《石河子大学》2017年硕士论文　论文类型：学位论文

【摘要】：干旱是我国主要的自然灾害之一,新疆属于干旱荒漠地带,气候干燥;同时新疆又是我国最大的棉花生产基地之一,而棉花属于耗水作物,农业用水危机越来越突出,培育抗旱转基因棉花显得尤为重要,可很大程度上缓解干旱对棉花的影响。抗旱转基因棉花的获得性抗性所引发的安全性问题则是其应用于生产和推广的前提。本研究以实验室通过花粉管通道法获得的11个转基因抗逆棉花株系和其3个亲本受体品种为材料,采用膜下滴灌,共5个不同灌水量处理:不灌溉(NI)、低量灌溉(LI)、中量灌溉(MI)、高量灌溉(HI)、正常灌溉(CK),其中正常灌水量略高于石河子棉区平均值。并对棉花农艺、生理、光合、荧光进行了测定,以期研究转基因棉花对水分的响应以及建立转基因棉花抗旱性评价体系用以评定其抗旱性和生存竞争力。并通过对连续耕作过转基因棉花和亲本棉花的根系土壤理化性质和微生物群落多样性进行分析,用以揭示抗旱转基因棉花对土壤微生物多样性造成的影响。为指导田间抗旱转基因棉花水分管理和环境安全性提供一定的理论依据和技术支持。主要研究结果如下:1.采用不灌溉(NI)和正常灌溉(CK)条件,通过主成分分析及隶属函数加权平均法(D值)并结合抗旱指数法对11个转基因抗逆棉花株系及其3种亲本受体的株高、红茎比、始果节位、果枝数、铃数、单铃重、单株铃重、籽棉产量、叶绿素、过氧化物酶(POD)、超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT)、可溶性蛋白等进行了抗旱综合性评价以鉴定其抗旱性强度。结果表明,在所有供试材料中,相对亲本受体型,转基因型的抗旱性均高于亲本。其中以25C-1和TH1-katG的抗旱性综合指标值最高,抗旱性最强,TH1-ACCD、TH1-35S-COR和24C-1为中间类型,TH1-HRD、TH1-SAD,672-SacB、672-RD-COR、672-RD-SAD和672-2300TF为不抗旱材料。证明外源抗逆基因的导入提高了其的抗旱性,对比亲本受体型其生存竞争力均有所提高。2.在5个不同灌水量处理下,对其中24C-1、25C-1两种转基因抗旱棉花株系进行研究,测定其在不同水分胁迫下棉花苗、蕾期农艺性状及花铃期不同阶段花(蕾)铃发育、叶片光合参数和叶绿素荧光参数等指标,研究转基因及其亲本受体棉花对水分胁迫响应机理的差异。结果表明,随着干旱胁迫程度的加重,棉花花(蕾)铃发育明显受到抑制,2个品种(系)的净光合速率(Pn)、PSII最大光化学效率(Fv/Fm)、光能利用率(LUE)、PSII光化学量子效率(φPSII)、光化学碎灭系数(qP)呈现下降趋势;而气孔限制值(Ls)和非光化学碎灭系数(NPQ)则呈现上升趋势,HI和CK下差异均不显著或差异较小。3.通过对25C-1及其对应亲本受体TH2棉花根际土壤采集,对根际土壤理化性质、作物产量进行连续3个种植季节及第三年进行的土壤微生物微生态多样性的测定,以期研究作物、土壤、土壤微生物三者间的动态变化关系。研究发现,在三年间转基因及其亲本受体在年与年及品种(系)间在PH值、全氮、速效磷和速效钾含量上均无显著差异,只在2014/2015、2014/2016的土壤有机质(OM)及土壤微生物微生态中Arthrobacter_oxydans和Lysobacter_soli的物种丰度上存在差异。结果表明,转基因及其亲本受体在连续种植季年间的土壤理化上无显著差异,而土壤有机质在年间出现显著差异,因此棉花秸秆还田可能是导致抗旱转基因型与亲本受体型根际微生态差异的根本原因。
[Abstract]:Drought is one of the major natural disasters in China, Xinjiang belongs to arid desert, dry climate; at the same time, Xinjiang is one of China's largest production base of cotton, and cotton to water consumption crops, agricultural water crisis has become increasingly prominent, the cultivation of drought resistance of transgenic cotton is particularly important, which can largely alleviate the effects of drought on cotton the drought resistance of transgenic cotton. The safety problems caused by resistance is applied to the production and promotion of the premise. In this study, the laboratory obtained by pollen tube pathway 11 transgenic cotton resistant strains and its receptor 3 parental varieties as materials, using drip irrigation, a total of 5 different irrigation the amount of processing: no irrigation (NI), low volume irrigation (LI), amount of irrigation (MI), a high amount of irrigation (HI), normal irrigation (CK), which is slightly higher than the normal amount of irrigation in Shihezi average. And on cotton agronomic, physiological, light Together, the fluorescence was measured, in order to study the response of transgenic cotton on the water and the establishment of Transgenic Cotton Drought Resistance evaluation system to evaluate the drought resistance and survival competitiveness. And through the root soil continuous cultivated transgenic cotton and cotton were analyzed the physicochemical properties and microbial community diversity, in order to reveal the impact of Drought Resistance Transgenic Cotton on soil microbial diversity caused. To provide theoretical basis and technical support for the guidance of drought resistant transgenic cotton field water management and environmental safety. The main results are as follows: 1. with no irrigation (NI) and normal irrigation (CK) conditions, through principal component analysis and membership function weighted average method (D value) combined with drought resistance index of 11 transgenic cotton resistant strains and 3 strains of high parental receptor, red stem, the first fruit node, branch number, boll number, boll weight per plant. Boll weight, seed yield, chlorophyll, peroxidase (POD), superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT), a comprehensive evaluation of drought resistance of soluble protein in identification of drought resistance and strength. The results showed that in all the tested materials, the relative by parents size, type of transgenic drought resistance. The drought resistance was higher than that of parent 25C-1 and TH1-katG index is the highest, the strongest drought resistance, TH1-ACCD, TH1-35S-COR and 24C-1 for TH1-HRD, TH1-SAD, intermediate type, 672-SacB, 672-RD-COR, 672-RD-SAD and 672-2300TF for drought resistant materials. That the exogenous genes into improved drought resistance the comparison of parents receiving the survival competitiveness increased.2. in 5 different irrigation treatments, the study of 24C-1,25C-1 two kinds of transgenic drought resistant cotton lines, measured in different water stress in cotton seedling, bud Different stages of agronomic traits and flowering stage (flower bud) Boll Development, photosynthetic parameters and chlorophyll fluorescence parameters, different response mechanism of transgenic cotton and its parental receptors to water stress. The results showed that with the aggravation of drought stress, Cotton Boll Development (bud) was significantly inhibited (2 varieties the Department of) the net photosynthetic rate (Pn), the maximum photochemical efficiency of PSII (Fv/Fm), the utilization rate of light energy (LUE), PSII photochemical efficiency (PSII), photochemical quenching coefficient (qP) decreased; while the stomatal limitation value (Ls) and non photochemical quenching coefficient (NPQ) is the rise, HI and CK had no significant difference or little difference of.3. on the 25C-1 and its corresponding receptor TH2 in cotton rhizosphere soil were collected on rhizosphere soil physicochemical properties, crop yield for 3 consecutive seasons and three years of planting soil microorganism ecological diversity The determination of crop, the soil, dynamic changes of soil microbial relationship between the three. The study found that in three years and their parents in transgenic receptor from year to year and varieties (lines) in pH, total nitrogen, available phosphorus and available potassium content were no significant differences in soil organic matter, only 2014/20152014/2016 the (OM) and soil microbial Arthrobacter_oxydans and Lysobacter_soli micro ecology of species abundance differences. The results showed that the transgenic and their receptors in the continuous planting season from soil had no significant difference, while soil organic matter has significant difference in years, so the cotton straw is the main reason for the drought resistant transgenic type and the parent body of rhizosphere differences.

【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S562;S423

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