Overexpression of the Suaeda salsa SsNHX1 gene confers enhanced salt and drought tolerance to transgenic Zea mays

Maize is one of the most important crops worldwide, but it suffers from salt stress when grown in saline-alkaline soil. There is therefore an urgent need to improve maize salt tolerance and crop yield. In this study, the SsNHX1 gene of Suaeda salsa, which encodes a vacuolar membrane Na⁺/H⁺ antiporter, was transformed into the maize inbred line 18-599 by Agrobacterium-mediated transformation. Transgenic maize plants overexpressing the SsNHX1 gene showed less growth retardation when treated with an increasing NaCl gradient of up to 1%, indicating enhanced salt tolerance. The improved salt tolerance of transgenic plants was also demonstrated by a significantly elevated seed germination rate (79%) and a reduction in seminal root length inhibition. Moreover, transgenic plants under salt stress exhibited less physiological damage. SsNHX1-overexpressing transgenic maize accumulated more Na⁺ and K⁺ than wild-type (WT) plants particularly in the leaves, resulting in a higher ratio of K⁺/Na⁺ in the leaves under salt stress. This result revealed that the improved salt tolerance of SsNHX1-overexpressing transgenic maize plants was likely attributed to SsNHX1-mediated localization of Na⁺ to vacuoles and subsequent maintenance of the cytosolic ionic balance. In addition, SsNHX1 overexpression also improved the drought tolerance of the transgenic maize plants, as rehydrated transgenic plants were restored to normal growth while WT plants did not grow normally after dehydration treatment. Therefore, based on our engineering approach, SsNHX1 represents a promising candidate gene for improving the salt and drought tolerance of maize and other crops.     

转LEA3基因水稻的抗性分析

对转LEA3基因水稻植株进行了抗渗透胁迫能力分析。结果显示在相同胁迫条件下,转基因水稻植株的苗高、根长、离体叶片保水率、叶绿素含量、可溶性糖含量、可溶性蛋白含量、SOD活性均高于未转化的对照植株,表明转LEA3基因水稻对盐分和干旱胁迫有较强的抗性。     

Overexpression of a maize SNF-related protein kinase gene,ZmSnRK2.11, reduces salt and drought tolerance in Arabidopsis

Sucrose non-fermenting-1 related protein kinase 2(Sn RK2) is a unique family of protein kinases associated with abiotic stress signal transduction in plants. In this study, a maize Sn RK2 gene Zm Sn RK2.11 was cloned and characterized. The results showed that Zm Sn RK2.11 is up-regulated by high-salinity and dehydration treatment, and it is expressed mainly in maize mature leaf. A transient expression assay using onion epidermal cells revealed that ZmS nR K2.11-GFP fusion proteins are localized to both the nucleus and cytoplasm. Overexpressing-Zm Sn RK2.11 in Arabidopsis resulted in salt and drought sensitivity phenotypes that exhibited an increased rate of water loss, reduced relative water content, delayed stoma closure, accumulated less free proline content and increased malondialdehyde(MDA) content relative to the phenotypes observed in wild-type(WT) control. Furthermore, overexpression of Zm Sn RK2.11 up-regulated the expression of the genes ABI1 and ABI2 and decreased the expression of DREB2 A and P5CS1. Taken together, our results suggest that Zm Sn RK2.11 is a possible negative regulator involved in the salt and drought stress signal transduction pathways in plants.     

异源表达玉米ZmC3H54基因增强水稻的耐旱性

CCCH锌指蛋白是一类重要的转录调控因子。从玉米中分离得到一个受干旱和ABA诱导表达的CCCH型锌指蛋白基因ZmC3H54,通过构建过量表达载体并转化水稻来进一步研究其功能。 与对照组相比,转基因植株在干旱胁迫处理下具有更高的相对含水量与存活率以及较低的相对电导率,表明过量表达ZmC3H54基因可以提高转基因水稻的耐旱性。此外,转基因水稻幼苗对外源ABA敏感性更高。以上结果表明玉米ZmC3H54基因可能是通过ABA信号通路调控植物对干旱的耐受性。     

过量表达AtNHXS1新基因显著提高水稻的耐盐性

以转基因和野生型水稻为材料,通过农杆菌介导法将AtNHXS1转到水稻植株中花11号中,分析在盐胁迫下Na+、K+含量的变化,对两者耐盐性进行比较,并对转基因株系进行分子鉴定和转录表达分析。结果表明:PCR初步鉴定得到了20个转基因株系,随机挑选2个PCR阳性株系进行Southern blot鉴定,确定AtNHXS1以单拷贝的形式成功插入到水稻基因组中。耐盐性分析表明,在盐胁迫条件下,转基因水稻植株的生长状况、干质量、鲜质量、Na+含量显著优于或高于野生型水稻植株;此外,300mmol/L NaCl处理下,转基因水稻植株能够正常存活,而野生型水稻5d内几乎全部死亡。将300mmol/L NaCl处理过的植株在无盐胁迫的条件下进行恢复生长试验,转基因植株10d内恢复正常,而野生型则不能。过量表达改组后的AtNHXS1新基因显著提高了水稻的耐盐性。     

Thellungiella halophila ThPIP1 gene enhances the tolerance of the transgenic rice to salt stress

Aquaporin proteins were demonstrated to play an important regulatory role in transporting water and other small molecules. To better understand physiological functions of aquaporins in extremophile plants, a novel ThPIP1 gene from the Thellungiella halophila was isolated and functionally characterized in the transgenic rice. Data showed that the ThPIP1 protein encoded 284 amino acids, and was identified to be located on the plasma membrane. The expression of ThPIP1 gene in the shoots and roots of T. halophila seedlings were induced by high salinity. The transgenic rice overexpressing ThPIP1 gene significantly increased plants tolerance to salt stress through the pathway regulating the osmotic potentials, accumulation of organic small molecules substances and the ratio of K+/Na+ in the plant cells. Moreover, split-ubiquitin yeast two-hybrid assay showed that Th PIP1 protein specifically interacted with ThPIP2 and a non-specific lipid-transfer protein 2, suggesting that ThPIP1 probably play a key role in responding to the reactions of multiple external stimulus and in participating in different physiological processes of plants exposed to salt stress.     

翦股颖AsEXP1基因的抗旱性分析

分析了不同匍匐翦股颖草坪草品种在干旱环境下AsEXP1基因的表达状况,发现AsEXP1基因的表达与草坪草品种的抗旱性呈显著正相关关系,亦即在耐旱草坪草品种中表达,而在不耐旱草坪草品种中不表达。对耐热但不耐旱草坪草品种‘PennA-4’的进一步检测分析发现,AsEXP1基因经高温诱导表达后,显著增强了‘PennA-4’品种的耐旱性。     

水稻苗期耐盐性基因SST分子标记的筛选与应用

以水稻耐盐突变体sst为对照,对67个水稻亲本及突变体sst与水稻品种华占的F2群体进行耐盐性筛选,并结合分子标记结果,分析水稻苗期耐盐性基因SST分子标记ID27093、ID27101、ID27118、INDEL1和INDEL2的特异性和准确性.结果表明,分子标记ID27093、ID27101、ID27118和INDEL1的扩增多态性较低;而分子标记INDEL2的扩增多态性较高,为共显性标记,具有较高的特异性,能区分耐盐供体sst与85.07%的供试亲本,且筛选准确率高.因此,分子标记INDEL2标记可以直接用于耐盐性基因SST分子标记辅助育种.     

利用选择回交导入群体进行粳稻优良恢复系抗旱性改良与QTL定位

以C418为轮回亲本,与供体亲本C71和早籼14分别进行杂交和2次回交,对BC2F2混合群体进行严格的干旱筛选,共得到16个抗旱导入系.在海南和北京的干旱和灌溉条件下对抗旱导入系BC2F5家系及其亲本进行抗旱性综合评价.结果表明,选择的导入系群体抗旱性平均表现与轮回亲本没有明显差异,不同组合在不同地点不同水分处理条件下的表现不尽一致,其中一些株系的抗旱性和产量均显著高于轮回亲本.产量构成因素中,单株有效穗数和每穗粒数与产量的关联密切,且最易受到干旱胁迫的影响,从而造成较大的产量损失.利用106个均匀分布的SSR标记对抗旱回交导入系进行基因型分析,通过导入频率的卡方检验共定位到16个抗旱QTLs(P＜0.0005),分布在除2、7号染色体外的10条染色体上,大部分导入的供体等位基因显示出正向加性效应.本研究结果说明,回交导入结合目标性状极端选择的策略改良水稻抗旱性并发掘和定位有利抗旱基因/QTL是可行的.     

小麦品种资源耐盐性鉴定

目的研究TaERECTA基因超表达后对小麦耐盐性的影响。方法以转TaERECTA基因小麦为材料,分别在芽期和苗期高盐胁迫,测定种子发芽势、发芽率、发芽指数和相对盐害指数,以及植株枯萎数、幼苗抗氧化酶活性和丙二醛(MDA)含量。结果芽期盐胁迫下,与野生型(WT)株系相比,TaERECTA转基因小麦种子发芽势、发芽率和发芽指数较高,相对盐害指数较低(P<0.05);苗期盐胁迫下,转基因小麦幼苗平均枯萎数较低,抗氧化酶活性较高,且SOD活性显著高于CK株系(P<0.05)。结论一定盐胁迫下,TaERECTA基因超表达可以增强小麦耐盐性,其作用机制可能是TaERECTA激活组织细胞SOD酶活性,其他抗氧化酶相互协同减轻盐胁迫伤害。转基因株系L10相对盐害指数较小,综合抗盐能力较强,可作为下一步深入研究TaERECTA调控小麦耐盐性功能的重要材料。     

Overexpression of MdMIPS1 enhances drought tolerance and water-use efficiency in apple

Myo-inositol and its derivatives play important roles in the tolerance of higher plants to abiotic stresses, and myo-inositol-1-phosphate synthase(MIPS) is the rate-limiting enzyme in myo-inositol biosynthesis. In this study, we found that increased myo-inositol biosynthesis enhanced drought tolerance in MdMIPS1-overexpressing apple lines under shortterm progressive drought stress. The effect of myo-inositol appeared to be mediated by the increased accumulation of osmoprotectants such as glucose...     

小麦水通道蛋白基因家族的全基因组鉴定及其在盐和干旱胁迫响应中的作用

水通道蛋白(AQPs)的功能是选择性地控制水和其他小分子通过细胞膜的流动,它们在植物的许多生理过程中都是至关重要的,包括非生物胁迫反应.小麦(Triticum aestivum L.)是全球重要的粮食作物,但干旱和盐胁迫是小麦生长和产量形成的重要影响因素.共鉴定得到了 127个非冗余的小麦水通道蛋白基因和4个可变剪接体.对TaAQP基因进行了 RNA-seq分析,揭示了小麦AQP基因的特异性表达模式.其中,TaTIPs和TaPIPs的表达高于TaNIPs和TaSIPs.qRT-PCR分析表明,小麦在干旱和盐胁迫条件下,TaNIP4;03_3D,TaTIP2;02b_7B,TaSIP2;02_4A,TaNIP3;03_6D 和 TaNIP2;04a_7D 等 AQPs 受到显著诱导并且有高表达量,表明它们参与了胁迫响应.这些结果为进一步探索TaAQPs基因在植物应对干旱胁迫和盐胁迫中的作用提供了新的思路.     

Identification of the DEAD-box RNA helicase family members in grapevine reveals that VviDEADRH25a confers tolerance to drought stress

Grapevine growing areas are increasingly affected by drought,which has greatly limited global wine production and quality.DEAD-box is one of the largest subfamilies of the RNA helicase family,and its members play key roles in the growth and development of plants and their stress responses.Previous studies have shown the potential of DEAD-box genes in the drought stress responses of Arabidopsis and tomato,rice,and other crop species.However,information about DEAD-box genes in grapevine remains li...     

胡杨PeAPY1和PeAPY2调控拟南芥耐盐机制研究

本文研究了胡杨apyrase基因(PeAPY1和PeAPY2)对植物耐盐性的影响。以PeAPY1和PeAPY2过表达拟南芥、拟南芥Atapy1和Atapy2突变体、野生型(WT)拟南芥及空载体(VC)为实验材料,研究盐胁迫条件下的植物根长、相对电导率、细胞活力、H₂O₂水平、eATP浓度、抗氧化酶活性的变化。研究结果显示:低盐浓度(50 mmol/L NaCl)对拟南芥各基因型的生长和生理生化指标没有显著影响;而高盐浓度(100 mmol/L NaCl)抑制了各株系的根长生长、细胞活力和抗氧化酶(超氧化物歧化酶/SOD、抗坏血酸过氧化酶/APX、过氧化氢酶/CAT)活性,却提高了相对电导率、H₂O₂水平和eATP浓度。但与Atapy突变体株系相比较,PeAPY过表达株系受高盐胁迫的影响较小。主要是由于PeAPY1/2的过表达提高了apyrase酶活,下调了eATP浓度及其诱发的活性氧水平,同时,过表达株系还通过保持抗氧化酶活性来抑制H₂O₂的水平,从而降低活性氧及其对膜脂的过氧化,保持了膜的稳定性、细胞活力和生长,最终提高了植物耐盐性。      

过表达玉米ZmSC1基因增强转基因拟南芥的耐盐性

旨在从玉米中克隆耐盐相关基因ZmSC1,分析其分子特征并在拟南芥中研究其耐盐性的生物学功能。以玉米B73为试验材料,克隆ZmSC1全长序列,与其他物种进行同源性比对,解析其盐诱导表达模式和亚细胞定位情况,将ZmSC1转入到拟南芥突变体atsc和野生型中,观察在盐处理下种子萌发率和主根根长情况,利用荧光定量PCR分析相关逆境（140 mmol·L^-1胁迫）基因的表达量。结果表明,ZmSC1基因全长为423 bp,编码141个氨基酸,ZmSC1和小麦、拟南芥中已知的TaSC1、AtSC1具有较高保守性。烟草细胞瞬时表达、玉米原生质体亚细胞定位研究表明ZmSC1定位于细胞膜中。生物学功能研究发现在140 mmol·L^-1的NaCl的盐处理下,相比较于拟南芥突变体,回补植株拟南芥的种子萌发率和主根根长得到了明显改善,这说明ZmSC1基因可以回补拟南芥同源基因AtSC1突变体植株在盐胁迫下的表型。ZmSC1基因过表达拟南芥植株的种子萌发率和主根根长也显著高于野生型植株。荧光定量PCR结果显示相较于野生型,在过表达植株中AtRD29A、AtSOS2、AtSOS3、AtCDPK1等胁迫相关基因的表达量也明显增强。研究结果表明TaSC1、AtSC1的同源基因ZmSC1对提高拟南芥的耐盐性具有重要作用。     

棉花耐盐碱性状的QTL定位

以感盐碱棉花品系新洋718和耐盐碱棉花品系苏研128构建的F2群体为材料,采用SSR分子标记构建了包含47个位点和16个连锁群的棉花分子遗传图谱。采用复合区间作图法对215个F2∶3家系的相对盐害率进行QTL定位,检测到6个与棉花耐盐碱性状相关的QTL,分别位于第3、5、7、7、15和19染色体上,分别解释3.23%、9.86%、5.77%、5.28%、11.31%和11.24%的表型变异。