转PvP5CS1基因拟南芥植株对干旱和盐胁迫的反应

为探索普通菜豆脯氨酸合成酶基因P5CS1在植物渗透胁迫中的作用,本研究应用农杆菌介导法,将PvP5CS1基因转入拟南芥,获得6株阳性转基因株系;通过检测转基因植株与野生型植株在干旱和盐胁迫下种子发芽率,幼苗脯氨酸含量、株系电导率、相对根长和成株死亡率,分析了PvP5CS1基因的表达对改善拟南芥抗渗透胁迫的效应。结果表明,在150 mmol L^-1 NaCl和150 mmol L^-1甘露醇渗透胁迫下,转基因植株平均相对发芽率分别是野生型的1.6倍和1.62倍;150、250 mmol L^-1甘露醇和150 mmol L^-1 NaCl处理下,转基因拟南芥植株平均脯氨酸含量分别是野生型的2.68、1.30和1.30倍;平均相对电导率分别是野生型植株的85%、77%和85%;平均相对根长分别是野生型植株的1.2、1.3和1.2倍;300 mmol L^-1 NaCl处理下,转基因植株的平均死亡率为42%,显著低于野生型(90%)(P<0.05);干旱胁迫下,转基因植株的平均死亡率为56%,显著低于野生型(70%)(P<0.05),说明PvP5CS1基因在拟南芥中的表达明显改善了转基因植株的抗旱性和耐盐性。     

蛋白激发子Hrip1基因在拟南芥中表达可提高植株的耐盐耐旱能力

Hrip1是从极细链格孢(Alternaria tenuissima)代谢物中分离的一种蛋白激发子。将蛋白激发子基因Hrip1转化到拟南芥,对5个T₁代转基因拟南芥株系进行分子检测, 证明Hrip1基因能够在拟南芥中转录和表达。转基因植株对盐和干旱胁迫的抗性显著增强, 75 mmol L⁻¹ NaCl和50 mmol L⁻¹甘露醇渗透胁迫2 d, 转基因植株种子平均相对发芽率为32.1%和77.9%, 分别比野生型的增加3.72倍和5.61倍; 150 mmol L⁻¹ NaCl和50 mmol L⁻¹甘露醇处理拟南芥幼苗7 d后, 转基因植株平均相对根长为81.79%和93.25%, 分别是野生型的1.53倍和1.34倍。3周龄的转基因植株在250 mmol L⁻¹ NaCl条件下胁迫20 d, 平均存活率为67%, 显著高于野生型(42%)(P<0.05); 干旱胁迫25 d后, 复水5 d转基因植株平均存活率为72%, 而野生型仅为44%。检测结果显示转基因植株叶片的抗氧化酶活性明显高于野生型, 用200 mmol L⁻¹ NaCl和200 mmol L⁻¹甘露醇处理24 h后, POD活性分别比野生型植株提高1.56倍和1.85倍, CAT活性分别比野生型植株提高1.64和1.86倍。说明蛋白激发子Hrip1基因在拟南芥中的表达能够改善和提高植株的耐盐抗旱能力。     

玉米胁迫诱导表达基因ZmSNAC1的功能分析

NAC转录因子是具有多种生物功能的植物特异转录调控因子,在植物生长发育、器官建成、激素调节和抵抗逆境等方面发挥着重要的作用。我们之前分离得到1个受干旱、盐、冷等非生物逆境胁迫和植物激素ABA诱导显著上调表达的玉米NAC家族成员ZmSNAC1,过表达转基因株系在苗期的耐脱水能力较野生型株系明显提高。在此基础上,本研究进一步对ZmSNAC1过表达转基因株系在植株生殖生长发育时期的抗旱性和耐盐性进行功能鉴定。结果表明,在干旱胁迫条件下,野生型株系相比转基因株系较存活率提高50%~52%,相对电导率降低17%~21%,叶绿素含量提高36%~47%,脯氨酸含量提高了17%~23%;在300mmol L^-1 NaCl胁迫条件下,转基因株系的存活率提高36~40%。ZmSNAC1可能作为一个正向调控因子在逆境胁迫信号转导过程中发挥重要作用。     

转betA基因的小黑杨花粉植株耐盐性分析

甘氨酸甜菜碱是植物细胞内一种重要的调渗物质,盐胁迫下,甘氨酸甜菜碱的积累可以保护细胞内蛋白质的结构和功能,降低细胞水势,从而增强植物自身的耐盐能力。从大肠杆菌中克隆的胆碱脱氢酶基因(betA)是甘氨酸甜菜碱合成的关键酶基因,该基因编码的胆碱脱氢酶(CDH)可将胆碱一步合成为甜菜碱。本实验室已将胆碱脱氢酶基因(betA)转入到小黑杨花粉植株基因组中,并最终获得了4个转基因株系。本研究以4个转betA基因株系(TB1、TB2、TB3、TB4)及非转基因对照为试材,在浓度为1.2%的NaCl盐胁迫下,测定其甜菜碱含量、超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性和丙二醛(MDA)含量,并调查试材的盐害情况,计算盐害指数,目的是为了研究转基因株系的耐盐效果,从中筛选出耐盐能力较强的转基因株系。试验结果表明,4个转基因株系的甜菜碱含量均高于非转基因对照;在1.2%NaCl胁迫下TB1、TB2、TB4的超氧化物歧化酶活性、过氧化物酶活性均高于非转基因对照,TB3低于对照;TB1、TB4的丙二醛含量低于对照,TB2、TB3与对照相近。进一步的盐害分析表明4个转基因株系中的TB1和TB4株系的盐害指数低于对照的42.1%和33.4%,TB2、TB3与对照无显著差异。综合各转基因株系的耐盐性及生理指标测定结果,4个转基因株系中,TB1、TB2的耐盐性明显优于对照,有希望用于盐碱地造林及推广。     

农杆菌介导的辽宁碱蓬胆碱单氧化酶基因CMO转化烟草提高抗氧化胁迫的研究

通过农杆菌介导的遗传转化,将来自辽宁碱蓬的甜菜碱合成关键基因－胆碱单氧化酶基因CMO 转入烟草,以提高烟草耐受氧化胁迫的能力,并对得到抗生素抗性植株进行了分子检测和百草枯诱导 的氧化胁迫。Southern 和Northern 检测表明CMO 已经整合到烟草基因组中,并得到了正确的表达,但 不同转基因株系中的拷贝数不同,株系T1、T2、T6 为单拷贝;株系T4、T5 为双拷贝;株系T3 为3 个拷 贝。在10 μM 和20 μM百草枯胁迫24 h 的情况下,转基因植株表现了较强的耐受氧化胁迫能力,各转 基因株系和野生型相比具有较低的丙二醛含量和较高的超氧化物歧化酶活性。这可能是转基因植物 中甜菜碱的积累诱导或保护了抗氧化酶的活性,使得转基因植株受到的氧化胁迫较低,转基因植株中 较低的电导率也证明了这一点。转CMO 基因烟草耐氧化胁迫能力的提高,说明CMO 基因可以进一步 在玉米、水稻等重要农作物中应用以提高其耐受干旱、低温等非生物胁迫的能力。     

强光高温同时作用下不同小麦品种的光合特性

以小麦品种藁城8901、豫麦49、郑麦9405、周18为试材,用强光(1 900 μmol m^-2 s^-1)、高温(35℃)同时处理材料3 h,研究了高光强和高温共同合胁迫对小麦旗叶光合作用和叶绿素荧光特性的影响。结果表明,正常生长条件下,郑麦9405和周18的光饱和速率、饱和光强均高于藁城8901和豫麦49,郑麦9405的表观量子效率也最高。强光高温处理使藁城8901、豫麦49和周18的光合速率(P_n)、蒸腾速率(T_r)、气孔导度(G_s)、胞间CO₂浓度(C_i)较对照大幅下降;郑麦9405的P_n、G_s也出现了下降,但降幅最小,P_n仍保持11.6 μmol CO₂ m^-2 s^-1,T_r较对照略有上升;4种基因型小麦的Ci未较对照明显变化;与对照相比,4个材料的最大光量子效率(F_v/F_m)、开放的光系统II反应中心的激发能捕获效率(F_v’/F_m’)、作用光下光系统II的实际量子效率(Ф_PSII)、光化学猝灭系数(q_P)均下降,而非光化学猝灭系数(NPQ)大幅上升;其中郑麦9405的各项荧光参数均较高。研究表明,小麦旗叶对强光高温的适应性存在品种间差异,郑麦9405的耐强光高温特性优于其他3品种;强光高温下较高的蒸腾速率和较大的NPQ可能是郑麦9405维持光合机构功能的重要原因。     

转入反义ScP5CS基因烟草植株对PEG和NaCl处理的反应

以转入甘蔗脯氨酸合成酶基因ScP5CS反义片段的转基因烟草与野生型烟草为材料,研究PEG6000与NaCl处理对转基因与野生型烟草植株生长及其T0代种子萌发的影响。结果表明,在17mmol/L NaCl与1%PEG6000胁迫下转入ScP5CS反义基因片段的烟草植株矮小,叶片容易发黄,根系发育迟缓,根长缩短;转基因烟草T0代种子在200mmol/L NaCl胁迫时其发芽率明显受到抑制,并随着盐胁迫处理时间达到第55天时,在50mmol/L NaCl胁迫下其叶片严重黄化,而野生型与转入空载体株系则相反。PEG6000处理对转基因烟草T0代种子萌发影响不大。可见,甘蔗ScP5CS基因在烟草中的反义表达,部分抑制了烟草P5CS基因的活力。     

转甜菜碱醛脱氢酶基因马铃薯的抗旱耐盐性

通过根癌农杆菌介导法将甜菜碱醛脱氢酶(BADH)基因导入马铃薯栽培品种甘农薯2号, 经PCR、Southern杂交和Northern杂交证明BADH基因已整合到马铃薯基因组中并在转基因植株中转录和表达。测定表明对照植株没有BADH酶活性, 各转化株系在胁迫前后BADH酶活性近似, 在2~11 U之间。BADH酶活性与叶片的相对电导率呈一定的负相关(y= –3.7738x+57.083, r=0.989^**)。在NaCl和PEG胁迫下, 转基因植株生长正常, 株高比对照提高0.41~1.00 cm, 单株重量比对照增加10%~35%, 说明外源BADH基因的导入提高了马铃薯植株对干旱和盐碱的抗性。     

转BADH基因玉米植株的获得及其耐盐性分析

采用超声波辅助花粉介导植物转基因方法, 将甜菜碱醛脱氢酶(BADH)基因导入玉米自交系郑58, 获得了耐盐性强的转基因玉米植株。经卡那霉素抗性初筛、PCR扩增、Southern blot杂交分析, 证明BADH基因已导入转化植株并整合到其基因组中。用不同浓度的NaCl溶液对T₂代转基因玉米植株与对照进行盐胁迫处理, 结果表明, 转BADH基因玉米植株表现出一定的抗逆性, 生长状况明显优于对照; 根据非转化苗对NaCl的反应以及生长状况, 确定250 mmol L^-1 NaCl溶液为玉米幼苗耐盐性筛选的适宜浓度; 依据此临界浓度下形态指标和生理生化指标的测定结果, 与对照相比, 转基因植株的株高提高10.94%~25.7%, 鲜重增加8.62%~18.2%, 干重增加9%~18.18%, 相对电导率降低37.21%~58.14%, 叶绿素含量增加15.89%~90.65%, 超氧化物歧化酶(SOD)活性提高64.92%~148.29%, 丙二醛(MDA)含量减少26.97%~48.05%。综上所述, 转入甜菜碱醛脱氢酶(BADH)基因提高了玉米的耐盐性。这是首例将BADH基因导入优良玉米自交系郑58的报道。超声波辅助花粉介导法是一种经济、高效、实用和无基因型依赖性的植物基因转化方法。     

转录因子BvM14-Dof3.4响应盐胁迫的功能研究

Dof(DNA binding with one finger)家族转录因子是植物特异的转录因子,在胁迫响应、种子发芽、氮同化、光合作用等多种生物学过程中发挥着重要作用。为了探究转录因子BvM14-Dof3.4在响应盐胁迫过程中的生物学功能,本研究以具有强耐盐特性的甜菜M14品系为试验材料,用花序浸染法将BvM14-Dof3.4基因在野生型拟南芥植株中异源表达,经150 mmol/L NaCl处理后发现,BvM14-Dof3.4基因的异源表达促进了盐胁迫下转基因拟南芥植株根的生长,提高了异源表达植株的鲜重和干重,与野生型拟南芥相比异源表达植株中K⁺/Na⁺比值增加1.3倍、甜菜碱含量增加1.1倍以及SOD和POD的酶活性分别上调1.3和1.2倍,从而减少了盐胁迫对异源表达拟南芥植株的损伤。这些结果表明了BvM14-Dof3.4基因响应盐胁迫,并且BvM14-Dof3.4基因的异源表达能够提高拟南芥植株的耐盐能力,该结果对甜菜M14品系优质基因资源的挖掘以及开展栽培甜菜抗逆性的遗传改良工作具有重要意义。     

Exogenous Glycinebetaine Enhances Grain Yield of Maize, Sorghum and Wheat Grown Under Two Supplementary Watering Regimes

Drought occurring at critical growth and developmental stages in cereals affects productivity by reducing biomass accumulation, grain set, and grain yield and quality. Maize (cv. SR-73), sorghum (cv. Trump), and wheat (cv. Spear) were established in drought-prone field conditions in Perth, Western Australia, in l994. The plants were then subjected to optimal and suboptimal supplementary watering regimes at growth stages that were sensitive to water availability. Glycinebetaine in aqueous solution was applied to leaves at three rates (2, 4 and 6 kg ha^?1 and a control) to establish whether its application could ameliorate the effects of drought on the yield of the crops. Above-ground biomass production was measured at the beginning and at termination of the watering regimes. Leaf tissue glycinebetaine concentrations were determined 1 and 3 weeks after application. At physiological maturity, grains from the crops were harvested and grain yield, number of grains m^?2 and single grain weight were recorded. Drought significantly reduced above-ground biomass production in maize (P = 0.047), but not in sorghum and wheat. Grain yield of maize, number of grains m^?2 of maize and sorghum, and single grain weight of sorghum were significantly depressed by drought. Foliar application of aqueous glycinebetaine marginally enhanced biomass production in the three crops and significantly increased grain yield of maize (P = 0.001) and sorghum (P = 0.003). It also resulted in more grains m^?2 of maize, sorghum and wheat (P = 0.001, 0.001 and 0.003, respectively), with interactions between water and glycinebetaine treatments for sorghum and wheat (P = 0.001 and 0.001. respectively). Residual tissue glycinebetaine levels remained high 3 weeks after application to the crops. The positive effects of glycinebetaine treatment appear to be linked to its physiological role as a plant osmoticum that improves drought tolerance. The results of these studies suggest that foliar application of glycinebetaine may be used to improve drought tolerance and economic yield of maize and sorghum, but not of wheat. Increased grain yield was associated with more grains m^?2 rather than greater single grain weight.     

Applications of Foliar Fertilizers Containing Glycinebetaine Improve Wheat Yields

Nitrogen availability and drought influence wheat (Triticum aestivum L.) grain yields in the semiarid and subhumid Pampas region of Argentina. The application of fertilizers containing osmoprotectants, such as glycinebetaine, to crop canopies might reduce crop losses caused by environmental stresses. The objectives of this study were (a) to determine the effects on wheat grain yields of the foliar application of a commercial fertilizer with glycinebetaine, and (b) to establish, under farmers’ field conditions, the relation between soil properties and the productivity of wheat crops treated with the same product. Two experiments were conducted in the north‐western part of the Buenos Aires province of Argentina on Typic Hapludolls and Entic Hapludolls. In the first, the treatments were N fertilization (0, 23 and 46 kg ha^?1) and the foliar application to the wheat crop, in the vegetative stages, of a foliar fertilizer containing N, P, Zn and glycinebetaine (0 and 2.5 l ha^?1). The second experiment was carried out on 10 farmers’ fields, for each of which the treatment was the foliar application to the wheat crop, in the vegetative stages, of the same foliar fertilizer (0 and 2.5 l ha^?1). In both experiments, the grain yield, the individual grain weight, the number of grains per spike and the spike and plant density at the physiological maturity of the crops were determined. Soil organic matter (SOM) and available P were determined in each of the 10 fields of the second experiment. Wheat grain yields were increased by N fertilization and glycinebetaine treatment. The number of grains per spike was higher in the treatments with glycinebetaine application. In the farmers’ fields, the grain yields of the treatments with the application of the product with glycinebetaine were, on average, 18 % higher than those of the non‐treated plots, with greater responses at sites with low SOM levels. We conclude that, in subhumid regions with a dry period between the tillering and flowering stages of wheat, the foliar application of foliar fertilizers containing glycinebetaine at the vegetative stages of crop development enhances grain yields by increasing the number of grains per spike. This response is independent of the SOM level or the N fertilization rate.     

根施甜菜碱对水分胁迫下小麦幼苗水分状况和抗氧化能力的改善作用

研究了外源根施甜菜碱对抗旱性不同的两个冬小麦品种幼苗抗旱性的影响。结果表明，水分胁迫会诱导小麦幼苗体内甜菜碱的积累，外源甜菜碱能提高体内甜菜碱含量，改善水分胁迫下两小麦品种的水分状况，提高其水势，膨压和相对含水量，这可能是通过提高小麦幼苗体内一些有机渗透调节物质（如可溶性糖和脯氨酸）含量进而增强其渗     

抗旱基因在小麦抗旱基因工程中的应用进展

干旱严重影响小麦的生长发育及产量,小麦抗旱育种是保障小麦生产的重要措施,利用基因工程技术提高小麦抗旱性是优于传统育种的有效途径。抗旱基因主要包括调节基因（蛋白激酶、蛋白酶和转录因子基因）和功能基因。目前,已证实的可提高小麦抗旱性的基因主要为转录因子基因CBF/DREB1、MYB、NAC（NAM、ATAF1、ATAF2和CUC2）、HD-Zip和WRKY等和功能基因LEA蛋白基因、甜菜碱合成酶基因和海藻糖合成酶基因等。本文从转录因子基因和功能基因2个方面概述国内外利用基因工程技术提高小麦抗旱性的研究进展,并对目前存在的问题进行分析,以期为小麦抗旱遗传改良及育种提供参考。     

河套灌区小麦耐盐性及其生态适宜区

以河套灌区小麦主栽品种永良4号为研究对象,选择20个农户地块定位观测,探讨不同土层土壤盐分含量对小麦产量及其构成因素的影响,研究小麦在该地区的耐盐性及其生态适宜区。结果表明,以小麦苗期0~10 cm土壤饱和浸提液电导率(ECe)作为根层盐分指标来评判小麦的耐盐性最具代表性,该土层对应的小麦耐盐指数为10.465;土壤盐分与小麦各产量构成因素之间均呈显著负相关,尤其与公顷穗数的相关性更强;根据小麦耐盐性分析,以小麦相对产量下降幅度≤10%、10%~25%、25%~50%和≥50%作为划分标准,将区域耕地划分为小麦生产最适宜区、适宜区、次适宜区和不适宜区,其对应的小麦苗期0~10 cm土壤ECe分别为≤4.972、4.972~6.747、6.747~9.196和≥9.196 dS m⁻¹。     

Improving resistance against terminal drought in bread wheat by exogenous application of proline and gamma‐aminobutyric acid

Scarcity of water is a severe constraint, which hinders the wheat productivity worldwide. However, foliage application of osmoprotectants may be useful in reducing the drought‐induced yield losses in wheat (Triticum aestivum L.). In this study, potential of foliage applied osmoprotectants (proline, gamma‐aminobutyric acid) in improving the performance of bread wheat against terminal drought was evaluated. Both proline and gamma‐aminobutyric acid (GABA) were foliage applied at 50, 100 and 150 mg/L at anthesis stage (BBCH‐identification code‐ 61), in two bread wheat cultivars viz. Mairaj‐2008 and BARS‐2009. After 1 week of foliage application of these osmoprotectants, drought was imposed by maintaining the pots at 35% water holding capacity. Imposition of drought caused significant reduction in the grain yield of both tested bread wheat cultivars; nonetheless, foliage applied osmoprotectants at either concentration improved the chlorophyll contents, accumulation of proline, glycinebetaine and total soluble phenolics and reduced the malondialdehyde contents, which resulted in better stay green, maintenance of grain weight and grain number under drought stress, thus resulting in better grain yield, water‐use efficiency and transpiration efficiency in both wheat cultivars. However, foliage applied proline at 150 mg/L, and GABA at 100 mg/L was most effective than other concentrations of these osmoprotectants. Performance of cultivar Mairaj‐2008 was quite better than cultivar BARS‐2009. In crux, foliar application of proline and GABA at pre‐optimized rate can be opted as a shotgun approach to improve the performance of wheat under terminal drought.     

Investigations on Heat Tolerance of Spring Wheat Varieties of Different Origin Under Growth Chamber Conditions

Winter wheat has replaced spring wheat to a large extent under the climatic conditions of Western and Central Europe. But spring wheat genotypes are still important under the warm climate in large parts of Africa and Central America, since spring wheat needs no vernalization. To reach optimal yield, genotypes must be well adapted to heat stress. In this study 11 spring wheat varieties of Egypt, Sudan, CIMMYT and Germany were exposed to high temperatures at ear emergence and beginning of anthesis. There were reductions in kernel yield and kernel number of the main ear for all genotypes, but the differences in reduction were remarkable. The German variety 'Naxos' was best adapted. Some varieties showed an increased number of florets per spikelet under heat treatment, but this does not prohibit a reduction in kernel number per spikelet.     

Improving Drought Tolerance by Exogenous Application of Glycinebetaine and Salicylic Acid in Sunflower

Water shortage is a severe threat to the sustainability of crop production. Exogenous application of glycinebetaine (GB) and salicylic acid (SA) has been found very effective in reducing the adverse affects of drought stress. This study was conducted to examine the possible role of exogenous GB and SA application in improving the yield of hybrid sunflower (Helianthus annuus L.) under different irrigation regimes. There were three levels of irrigation, viz. control (normal irrigations), water stress at vegetative stage (irrigation missing at vegetative stage) and water stress at flowering stage (irrigation missing at flowering stage). GB and SA were applied exogenously at 100 and 0.724 mm , respectively, each at the vegetative and at the flowering stage. Control plants did not receive application of GB and SA. Water stress reduced the head diameter, number of achene, 1000‐achene weight, achene yield and oil yield. Nevertheless, exogenous GB and SA application significantly improved these attributes under water stress. However, drought stress increased the free leaf proline and GB, and were further increased by exogenous application of GB and SA. However, exogenous GB application at the flowering stage was more effective than other treatments. Oil contents were also reduced under water stress; however, GB and SA application could not ameliorate the negative effect of water stress on achene oil contents. The effects of water stress and foliar application of GB were more pronounced when applied at the flowering stage than at the vegetative stage. Moreover, exogenous GB application was only beneficial under stress conditions.     

Seed Priming Enhances the Performance of Late Sown Wheat (Triticum aestivum L.) by Improving Chilling Tolerance

In rice–wheat systems, late sowing of wheat is the major reason of low yield. This yield reduction is principally due to lower and erratic germination, and poor crop establishment because of low temperature prevailing. The present study was conducted to explore the possibility of improving late sown wheat performance by seed priming techniques. Seed priming strategies were: on‐farm seed priming, hydropriming for 24 h, seed hardening for 12 h and osmohardening with KCl or CaCl₂ for 12 h. Seed priming improved emergence, stand establishment, tiller numbers, allometry, grain and straw yield, and harvest index. However, seed priming techniques did not affect plant height, number of spikelets, number of grains and 1000 grain weight. Osmohardening with CaCl₂ gave more grain and straw yield and harvest index compared with control and other priming treatments, followed by osmohardening with KCl and on‐farm seed priming. Improved yield was attributed principally to better stand establishment and improved number of fertile tillers. Seed priming techniques can be effectively used to improve the performance of late sown wheat.     

Wheat Breeding in a Water-stressed Environment. I. Delineation of Drought Tolerance and Susceptibility

To elucidate genotypic differences expressed through the grain yield of drought-stressed wheat, 21 commercial varieties and advanced lines were evaluated in the field under a range of soil water levels to induce varying degrees of drought stress. This paper presents data on grain yield and yield-based indices to indicate drought tolerance and drought susceptibility. AZS-4 was identified as drought tolerant and AZS-17 and ‘Pavon’ as drought susceptible. High gram yield under stress can be explained in terms of high yield potential, thus grain yield proved to be the best indicator of drought tolerance.