Involvement of a novel Arabidopsis phospholipase D, AtPLDdelta, in dehydration-inducible accumulation of phosphatidic acid in stress signalling

Phospholipid metabolism is involved in plant responses to drought and salinity stress. To investigate the role of phospholipase D (PLD) and its product phosphatidic acid (PtdOH) in stress signalling, we isolated a novel PLD cDNA, designated AtPLDdelta, by screening a cDNA library prepared from dehydrated Arabidopsis thaliana. The AtPLDdelta protein, of 868 amino acids, has a putative catalytic domain and a C2 domain that is involved in Ca2+/phospholipid binding. The AtPLDdelta mRNA accumulated in response to dehydration and high salt stress. Histochemical analysis showed that the AtPLDdelta gene is strongly expressed in the vascular tissues of cotyledons and leaves under dehydration stress conditions. Under normal growth conditions, AtPLDdelta was expressed in roots, leaves, stems and flowers but not in siliques. We showed that dehydration stimulates the accumulation of PtdOH. The accumulation of PtdOH in response to dehydration was significantly suppressed in AtPLDdelta antisense transgenic plants. These results suggest that AtPLDdelta may be involved in PtdOH accumulation in the dehydration stress response.     

小黑杨HD-Zip转录因子家族生物信息学及应答盐胁迫分析

HD-Zip转录因子基因是植物中特有的一类蛋白家族,在植物生长发育和逆境应答胁迫过程中发挥重要作用。HD-Zip转录因子基因是由高度保守的同源异型结构域（HD）和亮氨酸拉链域（LZ）结构域构成的特殊结构模型。杨树HD-Zip转录因子家族共有63个基因,可被分为HD-ZipⅠ、HD-ZipⅡ、HD-ZipⅢ和HD-ZipⅣ四个亚家族。本文利用RNA-Seq分析了盐胁迫条件下HD-Zip基因家族在小黑杨根、茎、叶等不同组织的基因表达差异,从转录组水平揭示其应答胁迫环境的分子机制,结果表明,盐胁迫下在叶中有25个HD-Zip基因下调表达,21个基因上调表达;茎中有42个基因下调表达,11个基因上调表达;根中有26个基因下调表达,24个基因上调表达。另外,本文根据拟南芥HD-Zip转录因子家族基因的已知功能,预测了杨树HD-Zip转录因子同源基因的功能,并利用生物信息学方法分析了杨树HD-Zip转录因子蛋白序列的保守结构域、氨基酸组成和理化性质等,为进一步研究杨树HD-Zip转录因子基因功能提供参考。     

XVSAP1 from Xerophyta viscosa improves osmotic-, salinity- and high-temperature-stress tolerance in Arabidopsis

XVSAP1, a gene isolated from a dehydrated Xerophyta viscosa cDNA library, was transformed into Arabidopsis thaliana by Ti plasmid-mediated transformation under the control of a cauliflower mosaic virus 35S promoter, a nos terminator and bar gene selection. Expression of XVSAP1 in Arabidopsis led to constitutive accumulation of the corresponding protein in the leaves. Transgenic Arabidopsis grown in tissue culture maintained higher growth rates during osmotic, high-salinity and high temperature stress, respectively. Non-transgenic plants had shorter roots, leaf expansion was inhibited and leaves were more chlorotic than those of the transgenic plants. This study demonstrates that XVSAP1 has a significant impact on dehydration, salinity and high-temperature stress tolerance in Arabidopsis.     

A poplar DRE-binding protein gene, PeDREB2L, is involved in regulation of defense response against abiotic stress

Dehydration responsive element (DRE)-binding proteins comprise a family of proteins that have important roles in abiotic stress processes. The present study aimed at elucidating the molecular basis of the stress tolerance mechanism in poplar. We isolated a gene, PeDREB2L, from the desert tree Populus euphratica Oliva. PeDREB2L contains a conserved AP2/ERF domain, a nuclear localization signal at the N-terminus and a serine-rich region at the C-terminus. PeDREB2L binds specifically to DRE elements and is expressed ubiquitously in all tissues, including roots, stems, and leaves. Expression of PeDREB2L in leaves increased under dehydration, salt, and abscisic acid stress treatments. Furthermore, PeDREB2L-GFP ectopically expressed in Arabidopsis gave strong green fluorescent protein signals in the nucleus of transgenic Arabidopsis under unstressed conditions and resulted in an improved tolerance against drought and freezing under stress conditions, while the Arabidopsis protein gene DREB2A-GFP used as a control only gave a weak fluorescent. We expected that the PeDREB2L might be useful in improving abiotic stress tolerance in transgenic plants.     

拟南芥ANAC055基因突变体对高盐和渗透胁迫的响应

高盐和渗透等非生物胁迫是影响农作物产量和品质的重要因素,非生物胁迫发生时,植物通过体内各类转录因子启动胁迫应答反应,进而降低非生物胁迫对植物的损伤。本研究筛选出植物特异性转录因子ANAC055编码基因的纯合T-DNA插入突变体SALK_152738,测序分析发现T-DNA插在ANAC055基因的3'UTR区域。实时荧光定量PCR结果表明叶中ANAC055基因表达量最高;与野生型相比,突变体叶、茎和花中ANAC055基因表达量分别下降了40%、50%和70%。高盐胁迫后,野生型和突变体叶中ANAC055基因表达量分别比对照上升了320%和55.4%;而渗透胁迫时,该基因叶中的表达量分别比对照下降了47.7%和56.3%;电子表达谱分析发现该基因根中的表达可受高盐和渗透等多种非生物胁迫的诱导表达。高盐和渗透胁迫时野生型和突变体幼根的生长均受到明显抑制,但高盐胁迫对突变体根生长的抑制作用比对野生型根生长的抑制作用更大。上述分析表明拟南芥ANAC055基因可受高盐和渗透等非生物胁迫的诱导表达,并且其在拟南芥幼根的生长发育过程中具有一定的作用,本研究有助于进一步明确其在非生物胁迫过程中的作用。     

Salt tolerance is unrelated to carbohydrate metabolism in cowpea cultivars

Under salinity stress, plants commonly accumulate carbohydrates for osmotic adjustment to balance the excess accumulated ions and to protect biomolecules. We selected two cowpea cultivars with contrasting response to salinity, Pitiúba (salt-tolerant) and TVu (salt-sensitive), to investigate whether the salt tolerance could be associated with changes in carbohydrate accumulation and metabolism in leaves and roots during a long-term experiment. Two salt treatments (0 and 75 mM NaCl) were applied to 10-day-old plants grown in nutrient solution for 24 days. Despite some changes in carbohydrate accumulation and carbohydrate metabolism enzymes induced by salt stress, no consistent alterations in carbohydrates could be found in leaves or roots in this study. Therefore, we suggest that tolerance to salt stress is largely unrelated to carbohydrate accumulation in cowpea.     

A comparative study of the early osmotic,ionic, redox and hormonal signaling response in leaves and roots of two halophytes and a glycophyte to salinity

Salt stress is one of the most important abiotic stress factors affecting plant growth and productivity in natural ecosystems. In this study, we aimed at determining possible differences between salt tolerant and salt sensitive species in early (within 72 h) salt stress response in leaves and roots. To this purpose, we subjected three Brassicaceae species, namely two halophytes—Cakile maritima and Thellungiella salsuginea—and a glycophyte—Arabidopsis thaliana— to short-term salt stress (400 mM NaCl). The results indicate that the halophytes showed a differential osmotic and ionic response together with an early and transient oxidative burst, which was characterized by enhanced hydrogen peroxide levels and subsequent activation of antioxidant defenses in both leaves and roots. In addition, the halophytes displayed enhanced accumulation of abscisic acid, jasmonic acid (JA) and ACC (aminocyclopropane-1-carboxylic acid, the precursor of ethylene) in leaves and roots, as compared to A. thaliana under salt stress. Moreover, the halophytes showed enhanced expression of ethylene response factor1 (ERF1), the convergence node of the JA and ethylene signaling pathways in both leaves and roots upon exposure to salt stress. In conclusion, we show that the halophytes C. maritima and T. salsuginea experience an early oxidative burst, improved antioxidant defenses and hormonal response not only in leaves but also in roots, in comparison to the glycophyte A. thaliana. This differential signaling response converging, at least in part, into increased ERF1 expression in both above- and underground tissues seems to underlay, at least in part, the enhanced tolerance of the two studied halophytes to salt stress.     

6-苄（基）腺嘌呤和抗坏血酸对渗透胁迫时杨树光合作用光抑制的影响

研究了6-BA和A5A对渗透胁迫时杨树幼苗叶片光合作用光抑制和活性氧代谢的影响．结果表明,渗透胁迫时杨树叶片净光合速率(Pn)和表观量子效率(AQY)降低,光合作用光抑制加剧,超氧化物歧化酶(SOD)活性升高,抗坏血酸过氧化物酶(APX)活性降低,O2产生加快,H2O2和膜脂过氧化产物丙二醛(MDA)含量升高．6-BA和A5A预处理使胁迫时叶片SOD和APx活性升高。O2生成减少。H2O22和MDA含量降低,同时缓解了光合作用的光抑制．相关分析表明,杨树叶片活性氧水平和MDA含量与Pn和AQY呈负相关．胁迫时杨树叶片活性氧的积累与光合作用光抑制有一定关系,6-BA和A5A对光抑制的缓解作用与其对活性氧清除系统的促进作用有关。     

Induction of salt and osmotic stress tolerance by overexpression of an intracellular vesicle trafficking protein AtRab7 (AtRabG3e)

Adaptation to stress requires removal of existing molecules from various cellular compartments and replacing them with new ones. The transport of materials to and from the specific compartments involved in the recycling and deposition of macromolecules is carried out by an intracellular vesicle trafficking system. Here, we report the isolation of a vesicle trafficking-regulating gene, AtRabG3e (formerly AtRab7), from Arabidopsis. The gene was induced during programmed cell death after treatment of intact leaves with superoxide and salicylic acid or infection with necrogenic pathogens. Transgenic plants that expressed the AtRabG3e gene under the constitutive 35S promoter from cauliflower mosaic virus exhibited accelerated endocytosis in roots, leaves, and protoplasts. The transgenic plants accumulated sodium in the vacuoles and had higher amounts of sodium in the shoots. The transgenic plants also showed increased tolerance to salt and osmotic stresses and reduced accumulation of reactive oxygen species during salt stress. These results imply that vesicle trafficking plays an important role in plant adaptation to stress, beyond the housekeeping function in intracellular vesicle trafficking.     

5个毛果杨PtrZFP基因的鉴定和表达分析

ZFP转录因子是植物中的一类具有指环结构域的转录因子。从毛果杨中鉴定出5条ZFP基因（命名为PtrZFP1-5）,对其特性和表达模式进行了分析,以期初步了解这些基因是否能对胁迫做出应答。对PtrZFP1-5基因进行生物学分析,进一步利用qRT-PCR技术分析NaCl、PEG₆₀₀₀和ABA胁迫处理后毛果杨根、茎和叶中5条基因的表达情况。PtrZFP1-5基因编码蛋白氨基酸残基数为258~338 aa,编码蛋白的分子量为27.7~37.3 kDa,理论等电点为4.87~8.61,5个基因不均等的分布在毛果杨基因组的3条染色体上。qRT-PCR结果显示,0.2 mol·L^-1 NaCl、15%（w/v）PEG6000和100 μmol·L^-1 ABA胁迫处理后,5个PtrZFP基因在毛果杨根、茎和叶中的表达模式明显不同。PtrZFP1基因在3种胁迫后毛果杨中均被明显的上调表达;PtrZFP2基因在盐、渗透和ABA胁迫处理后,叶中的表达都明显被抑制;PtrZFP3基因受到干旱胁迫时在根中的响应最为明显;而叶和茎中,表达量在大部分胁迫的大部分时间点无明显改变。PtrZFP4基因也能在根和茎中对干旱胁迫做出明显应答。PtrZFP5基因在经受盐和ABA胁迫后,在叶中的表达受到明显抑制。PtrZFP1-5这5个基因至少能在一种器官中对一种胁迫处理做出应答,但参与的胁迫应答类型和机制可能不同。