非生物胁迫对花生叶片AhNCED1蛋白表达的影响

AhNCED1(9-cis-epoxycarotenoid dioxygenase 1 in Arachis hypogaea L.)是调控花生ABA(abscisic acid)生物合成的关键限速酶.利用Western blotting研究不同非生物胁迫对花生叶片AhNCED1蛋白表达的影响.结果表明渗透胁迫引起叶片AhNCED1蛋白表达快速(1 h)增加,ABA含量不断升高;NaCl处理后叶片AhNCED1蛋白在胁迫中期(7 h)表达增加,胁迫10 h时ABA含量显著升高;低温处理后叶片AhNCED1蛋白在胁迫后期(10 h)表达增加;高温胁迫后AhNCED1蛋白表达无明显变化.说明花生在感受(高渗、高盐、低温)等非生物胁迫后可能通过激活AhNCED1蛋白的表达,促使花生内源ABA水平升高以适应外界环境.     

综合隶属函数法评价花生品种抗旱性与AhNCED1基因表达的关系

利用综合隶属函数法评价不同产地8个花生品种幼苗的抗旱能力, 同时分析各品种花生在模拟干旱处理2小时和12小时后叶片AhNCED1基因表达量的变化, 探讨花生品种抗旱性与叶片AhNCED1基因表达的关系, 建立花生(Arachis hypogaea)幼苗时期通过AhNCED1基因表达变化检测花生抗旱性等级的方法。结果显示, 干旱处理2小时, 各品种叶片AhNCED1基因表达量迅速增加, 与处理前相比差异极显著; 干旱处理12小时, 各品种花生叶片AhNCED1基因表达量均降低, 但仍均高于处理前水平。综合隶属函数值定量反映各品种抗旱性强弱顺序为: 花育24号>福花13号>粤油7号>中花15号>中花16号>航花2号>福花9号>北海1号。干旱处理2小时各品种花生叶片AhNCED1表达量变化大小依次为: 福花13号>花育24号>中花16号>中花15号>航花2号>粤油7号>北海1号>福花9号。其中, 抗旱性强的品种为花育24号和福花13号, 抗旱性弱的品种为福花9号和北海1号, 其余4个品种为中等抗旱。该抗旱性评价结果与这些品种在生产上的表现一致, 干旱胁迫2小时花生品种叶片AhNCED1基因的表达量变化与其抗旱性一致。因此, AhNCED1基因表达量可作为苗期花生品种抗旱性等级的量化指标之一。     

不同花生品种响应干旱胁迫后叶片内ABA与AhNCED1的分布

以粤油7号和汕优523两个不同抗旱性品种为材料,研究响应干旱胁迫后叶片ABA(abscisic acid,脱落酸)和AhNCED1(Arachis hypogaea nine-cis-epoxycarotenoid dioxygenase)的分布以及含量变化。结果表明,两种花生品种响应干旱胁迫后叶片的维管组织中ABA分布增强且含量增加,AhNCED1蛋白分布也增强;且在水分胁迫初期粤油7号花生AhNCED1蛋白分布强于汕优523,其体内ABA分布水平也高于汕优523;经ABA生物合成抑制剂Naproxen处理后,两种花生叶片ABA分布减弱,但粤油7号叶片维管组织中ABA分布水平仍高于汕优523。结果表明维管组织是干旱胁迫下花生叶片中ABA和AhNCED1分布的主要区域,且粤油7号花生抗旱性强可能与其体内AhNCED1和ABA的分布量较高有关。     

花生响应水分胁迫过程中叶片AhCYP707A1蛋白和ABA分布的变化

CYP707A蛋白是高等植物内源ABA代谢主要途径的关键酶,在水分胁迫过程中花生植株Ah CYP707A1蛋白和ABA分布与水平的变化并不清楚。研究结果表明,水分胁迫初期,花生叶片Ah CYP707A1蛋白表达均先被强烈抑制,促进ABA积累,随后蛋白表达回升,参与调节ABA稳态;叶片Ah CYP707A1蛋白以及ABA主要分布在叶片维管组织中。水分胁迫下,经CYP707A蛋白抑制剂处理后,花生叶片ABA分布和含量均有提高。抗旱花生品种‘粤油7’在水分胁迫下,Ah CYP707A1蛋白的表达以及分布均强于敏旱品种‘汕优523’,表明ABA代谢更为活跃。     

转AhNCED1基因拟南芥的形态特征和抗旱性研究

目的：探讨外源基因花生NCED1（AhNCED1）对拟南芥的形态特征和抗旱能力的影响。方法：观察转AhNCED1拟南芥的表型特征以及在干旱条件下的变化,利用扫描电镜观察植株叶上表面细胞和气孔大小。结果：与野生型相比,AhNCED1转基因植株矮壮,株型紧凑,分蘖和分枝数减少,幼苗根发育好。在干旱条件下转基因型植株叶大色绿,气孔开度减小,表皮细胞排列紧密。结论：转基因型植株在干旱条件下的表型特征与抗旱能力表明转基因拟南芥抗旱能力高于野生型。     

水分胁迫对不同基因型小麦幼芽蛋白质表达和某些生理特性的影响

以抗旱小麦品种长武134和不抗旱小麦品种郑引1号为试材,采用-1.2 MPa PEG 6000处理种子,研究不同水分条件下小麦幼芽中蛋白质表达和部分生理特性的变化.聚丙烯酰胺凝胶电泳结果表明：水分胁迫可诱导抗旱品种幼芽产生分子量约39.5 kDa和23.0 kDa两种新蛋白亚基,不抗旱品种则无新亚基产生;在正常供水条件下,随着生育期延长,两品种中分子量为48.5 kDa的蛋白亚基表达量逐渐增强,因其对水分敏感且属于新发现蛋白,初步命名为水敏感蛋白.生理特性测定结果表明,水分胁迫条件下长武134的根芽比和相对含水量均高于郑引1号,而细胞膜相对透性和丙二醛含量则低于郑引1号.     

水分胁迫下高粱叶片的渗透调节与延伸生长的关系

高粱抗旱品种3197B比不抗旱品种三尺三在水分胁迫条件下ψ_S下降低。在相同ψ_S时,3197B相对含水量高于三尺三。水分胁迫期间,3197B能始终维持比三尺三较高的ψ_P。在中度和严重水分胁迫时,3197B几种渗透物质积累均高于三尺三,其中可溶性糖和K~ 对渗透调节贡献最大。水分胁迫下,3197B正展开叶渗透调节能力较强,ψ_P维持较高,临界膨压低,叶片扩张性能小、故生长速率随ψ_W下降较慢。     

AhAO2转基因拟南芥植株抗旱生理分析

目的:研究AhAO2基因对拟南芥抗旱生理的影响.方法:以转AhAO2基因拟南芥(AO2 -1 -4、AO2 -3 -7、AO2 -8 -1)为实验材料,通过植物根长、相对含水量、气孔开度和抗氧化酶比活力指标的综合评定,分析超表达AhAO2基因对拟南芥抗旱生理的影响.结果:在PEG胁迫下,AO2 -1 -4、AO2 -3 -7、AO2 -8 -1植株体内相对含水量分别比野生型高2.1％、1.3％和1.6％;超表达植株AO2 -1 -4、AO2 -3 -7、AO2 -8 -1抗氧化酶POD酶比活力较野生型提高73.1％、66.2％和74.4％,SOD酶比活力较野生型提高64.6％、80.5％和43.1％.而野生型的气孔开度在正常生长和PEG胁迫的条件下均低于超表达拟南芥株系.AhAO2转基因植株可能通过提高抗氧化酶活性提升了抗旱能力.     

干旱胁迫下不同抗旱性小麦叶片内源激素含量的变化与抗旱力强弱的关系

以抗旱性强的小麦品种昌乐5号和抗旱性弱的鲁麦5号的幼苗为材料,研究了随着干旱胁迫的加剧,小麦叶片相对含水量、气孔导度和内源激素水平的变化．结果表明,叶片的气孔导度和细胞激动素与脱落酸含量的比值（CTKs/ABA）呈较强的线性正相关关系而与叶片RWC的相关关系较弱,说明气孔导度受CTKs/ABA调控,而不是受叶片RWC的调控．在不同强度的干旱胁迫下我们可以利用CTKs/ABA的变化判断品种抗旱性大小：①抗旱力强的小麦品种叶片的平均CTKs/ABA值较高,而弱者较低,说明强者内源促进型激素含量相对较高,从而在干旱胁迫下保持较高的生活力．②抗旱力强的小麦品种叶片CTKs/ABA与干旱处理天数呈二次负相关关系,而弱者是线性负相关关系,说明强者具有较强的抗逆缓冲能力．③抗旱力强的小麦品种成熟叶（第二叶）CTKs／ABA下降快,新叶（第三叶）下降慢,而弱者反之,说明抗旱性强者可能存在着较强的从成熟叶向新叶的物质运输从而具有自我保护性调节机制．     

不同抗旱性花生品种结荚期叶片生理特性

以12个花生品种为试验材料,在人工控水条件下,于结荚期设置0~80 cm土层相对含水量为70%和50% 2个处理,研究与花生抗旱性相关的叶片生理生化性状及不同品种抗旱的叶片机制.结果表明： 利用产量抗旱系数可将12个花生品种的抗旱性分为强、中、弱3级,抗旱性强的品种有A596、山花11号、如皋西洋生,中度抗旱品种有花育20、农大818、海花1号、山花9号和79266,抗旱性弱的品种有ICG6848、白沙1016、花17和蓬莱一窝猴.A596、山花11号、如皋西洋生的抗旱机制是具有较强的叶片抗氧化保护能力、较高的PSⅡ活性及光合速率(P_n).海花1号的叶片抗氧化保护能力较强,山花9号的PSⅡ活性有显著优势.相关分析表明,叶片P_n、气孔限制值(L_s)、最大光化学效率(F_v/F_m)、光化学猝灭系数(q_P)、丙二醛(MDA)含量、相对电导率和超氧化物歧化酶(SOD)活性与花生品种的抗旱系数有显著的相关性,是花生结荚期的重要抗旱性状.SOD活性的抗旱级别需在干旱胁迫下鉴定,其他性状可在正常灌水条件下鉴定.山花11号和79266可分别作为花生强、弱抗旱性鉴定的标准品种,山花11号可作为花生叶片优异抗旱性状鉴定的标准品种.     

Light-Induced Stomatal Opening Is Affected by the Guard Cell Protein Kinase APK1b

Guard cells allow land plants to survive under restricted or fluctuating water availability. They control the exchange of gases between the external environment and the interior of the plant by regulating the aperture of stomatal pores in response to environmental stimuli such as light intensity, and are important regulators of plant productivity. Their turgor driven movements are under the control of a signalling network that is not yet fully characterised. A reporter gene fusion confirmed that the Arabidopsis APK1b protein kinase gene is predominantly expressed in guard cells. Infrared gas analysis and stomatal aperture measurements indicated that plants lacking APK1b are impaired in their ability to open their stomata on exposure to light, but retain the ability to adjust their stomatal apertures in response to darkness, abscisic acid or lack of carbon dioxide. Stomatal opening was not specifically impaired in response to either red or blue light as both of these stimuli caused some increase in stomatal conductance. Consistent with the reduction in maximum stomatal conductance, the relative water content of plants lacking APK1b was significantly increased under both well-watered and drought conditions. We conclude that APK1b is required for full stomatal opening in the light but is not required for stomatal closure.     

Water status of drought-resistant and drought-sensitive sorghum treated with ethephon

The effects of ethephon on stomatal resistance, water potential, osmotic potential, turgor potential, and ethylene production were determined on leaves of a drought-resistant (KS 65) and a drought-sensitive (IA 25) genotype of sorghum [Sorghum bicolor (L.) Moench] grown under wellwatered or drought-stressed conditions. With both sufficient and limited water supply, ethephon had no effect on the adaxial, abaxial, or total stomatal resistance of either genotype. For both water treatments, the adaxial stomatal resistance of the drought-sensitive genotype was higher than that of the drought-resistant genotype. Ethephon increased the amount of ethylene produced by the plants under both levels of water. For plants with sufficient water, water potentials of both genotypes were lowered by ethephon. Ethephon had no effect on the water potentials under drought or on the osmotic potentials under either water regime. With drought, the turgor potential of the drought-sensitive genotype, but not that of the drought-resistant, was increased by ethephon.     

The higher expression and distribution of 9-cis-epoxycarotenoid dioxygenase1 (AhNCED1) from Arachis hyopgaea L. contribute to tolerance to water stress in a drought-tolerant cultivar

The 9-cis-epoxycarotenoid dioxygenase (NCED) is thought to be the rate-limiting enzyme in the abscisic acid (ABA) biosynthetic pathway. In this study, transient expression of AhNCED1 and ABA distribution were detected in the vascular cambium of a drought-tolerant peanut cultivar (Yueyou 7) under a water stress treatment. It caused increases in ABA content in this region. The synthesis of ABA and AhNCED1 in the leaves of Yueyou 7 took place more quickly than in the control cultivar (Shanyou 523). Furthermore, AhNCED1 mRNA and proteins were induced in Yueyou 7 than in Shanyou 523, coinciding with greater ABA accumulation. During the seedling, blooming, and fruiting stages, AhNCED1 protein expression was higher in Yueyou 7 than in Shanyou 523, and it was induced more quickly when the plants were under water stress. These data suggest that the drought-tolerant cultivar can synthesize and distribute ABA more rapidly than does the control cultivar because of a high level of AhNCED1 expression, which then modulates physiological responses under water stress conditions.     

Transcriptional profiles of primary metabolism and signal transduction-related genes in response to water stress in field-grown sunflower genotypes using a thematic cDNA microarray

A sunflower cDNA microarray containing about 800 clones covering major metabolic and signal transduction pathways was used to study gene expression profiles in leaves and embryos of drought-tolerant and -sensitive genotypes subjected to water-deficit stress under field conditions. Using two-step ANOVA normalization and analysis models, we identified 409 differentially expressed genes among genotypes, water treatment and organs. The majority of the cDNA clones differentially expressed under water stress was found to display opposite gene expression profiles in drought-tolerant genotype compared to drought-sensitive genotype. These dissimilarities suggest that the difference between tolerant and non-tolerant plants seems to be associated with changes in qualitative but not quantitative mRNA expression. Comparing leaves and embryos, 82 cDNA clones showing organ-specific variation in gene expression levels were identified in response to water stress across genotypes. Genes related to amino acids and carbohydrates metabolisms, and signal transduction were induced in embryos and repressed in leaves; suggesting that vegetative and reproductive organs respond differentially to water stress. Adaptive mechanisms controlling water deficit tolerance are proposed and discussed.     

Water status of drought-resistant and drought-sensitive sorghum treated with ethephon

The effects of ethephon on stomatal resistance, water potential, osmotic potential, turgor potential, and ethylene production were determined on leaves of a drought-resistant (KS 65) and a drought-sensitive (IA 25) genotype of sorghum [Sorghum bicolor (L.) Moench] grown under wellwatered or drought-stressed conditions. With both sufficient and limited water supply, ethephon had no effect on the adaxial, abaxial, or total stomatal resistance of either genotype. For both water treatments, the adaxial stomatal resistance of the drought-sensitive genotype was higher than that of the drought-resistant genotype. Ethephon increased the amount of ethylene produced by the plants under both levels of water. For plants with sufficient water, water potentials of both genotypes were lowered by ethephon. Ethephon had no effect on the water potentials under drought or on the osmotic potentials under either water regime. With drought, the turgor potential of the drought-sensitive genotype, but not that of the drought-resistant, was increased by ethephon.Contribution no. 90-147-J of the Journal Series of the Kansas Agricultural Experiment Station, Manhattan, Kansas, USA.