Drought Priming at Vegetative Growth Stage Enhances Nitrogen‐Use Efficiency Under Post‐Anthesis Drought and Heat Stress in Wheat

To study the effects of early drought priming at 5th‐leaf stage on grain yield and nitrogen‐use efficiency in wheat (Triticum aestivum L.) under post‐anthesis drought and heat stress, wheat plants were first exposed to moderate drought stress (drought priming; that is, the leaf water potential reached ca. ?0.9 MP a) at the 5th‐leaf stage for 11 days, and leaf water relations and gas exchange rates, grain yield and yield components, and agronomic nitrogen‐use efficiency (ANUE ) of the primed and non‐primed plants under post‐anthesis drought and heat stress were investigated. Compared with the non‐primed plants, the drought‐primed plants possessed higher leaf water potential and chlorophyll content, and consequently a higher photosynthetic rate during post‐anthesis drought and heat stress. Drought priming also resulted in higher grain yield and ANUE in wheat under post‐anthesis drought and heat stress. Drought priming at vegetative stage improves carbon assimilation and ANUE under post‐anthesis drought and heat stress and their combination in wheat, which might be used as a field management tool to enhance stress tolerance of wheat crops to multiple abiotic stresses in a future drier and warmer climate.     

Improved Cold and Drought Tolerance of Doubled Haploid Maize Plants Selected for Resistance to Prooxidant tert‐Butyl Hydroperoxide

To improve the abiotic stress tolerance of maize (Zea mays L.), doubled haploid (DH) plants were produced by in vitro selection of microspores exposed to tert‐butyl hydroperoxide (t‐BuOOH) as a powerful prooxidant This study investigated the tolerance of the progenies of t‐BuOOH‐selected DH lines to oxidative stress, cold and drought in controlled environment pot experiments by analyses of photosynthetic electron transport and CO₂ assimilation processes, chlorophyll bleaching and lipid peroxidation of leaves. Our results demonstrated that the t‐BuOOH‐selected DH plants exhibited enhanced tolerance not only to oxidative stress‐induced by t‐BuOOH but also to cold and drought stresses. In addition, they showed elevated activities of antioxidant enzymes such as superoxide dismutase, ascorbate peroxidase, catalase, glutathione reductase and glutathione S‐transferase when compared with the DH lines derived from microspores that were not exposed to t‐BuOOH and to the original hybrid plants. The results suggest that the simultaneous up‐regulation of several antioxidant enzymes may contribute to the oxidative and cold stress tolerance of the t‐BuOOH‐selected DH lines, and that the in vitro microspore selection represents a potential way to improve abiotic stress tolerance in maize.     

Genome‐Wide Identification of MicroRNAs Responsive to High Temperature in Rice (Oryza sativa) by High‐Throughput Deep Sequencing

MicroRNAs (miRNAs) are known to play important roles in plant growth and stress response. Heat stress is a severe abiotic stresses by adversely affecting plant growth and yield. To identify heat‐responsive miRNAs at the genome‐wide level in rice (Oryza sativa), we constructed two small RNA libraries from young panicles treated or not with heat conditions. Ion torrent sequencing of the two libraries identified 294 known miRNAs and 539 novel miRNAs. Differential expression analysis showed that 26 miRNAs were downregulated and 21 miRNAs were upregulated in response to heat stress. Among them, five heat‐responsive miRNAs, including miR162b, miR529a‐p5, PC‐5P‐62245‐9, miR171b and miR169n, were validated by quantitative real‐time polymerase chain reaction. A total of 44 target genes of the differentially expressed miRNAs were predicted. These target genes are most significantly overrepresented in the cell growth process. The results demonstrated that rice miRNAs play critical roles in the heat stress response. This study opens up a new avenue for understanding the regulatory mechanisms of miRNAs involvement in the heat stress response in rice.     

QTL mapping for heading date,leaf area and chlorophyll content under cold and drought stress in two related recombinant inbred line populations (Japonica rice) and meta‐analysis

Rice is highly susceptible to drought and cold. The goal of this study was to identify the QTLs affecting the rice heading date (HD), leaf area (LA) and chlorophyll content (CC) under cold and drought stress. A total of twenty‐nine and thirty‐eight additive QTLs were detected from two crosses of ‘Dongnong422’/‘Kongyu131’ and ‘Xiaobaijingzi’/‘Kongyu131’, respectively. qHD1‐7‐1, qHD1‐7‐2, qHD1‐2‐1, qLA1‐7‐1, qLA1‐6‐3 and qCC1‐7‐1 show adaptive effects under cold treatment, while qHD2‐2‐3, qHD2‐2‐2, qLA2‐7‐3 and qCC2‐5‐1 were important for explaining the genetic mechanism during drought tolerance. Additionally, nine and five additive × environment interaction QTLs were detected for two RILs, respectively. RIL26 and RIL73 were two lines that are associated with cold and drought for HD. 339 QTLs related to HD, CC and LA of rice from database and our research were projected onto the genetic map. Nine cloned genes and nineteen homologous candidate genes related to HD, CC, cold tolerance and drought tolerance were mapped by meta‐analysis. These results lay the foundation for the fine mapping of QTLs related to HD, LA and CC for marker‐assisted selection.     

Integrated breeding approaches for improving drought and heat adaptation in chickpea (Cicer arietinum L.)

Chickpea (Cicer arietinum L.) is a dry season food legume largely grown on residual soil moisture after the rainy season. The crop often experiences moisture stress towards end of the crop season (terminal drought). The crop may also face heat stress at the reproductive stage if sowing is delayed. The breeding approaches for improving adaptation to these stresses include the development of varieties with early maturity and enhanced abiotic stress tolerance. Several varieties with improved drought tolerance have been developed by selecting for grain yield under moisture stress conditions. Similarly, selection for pod set in the crop subjected to heat stress during reproductive stage has helped in the development of heat‐tolerant varieties. A genomic region, called QTL‐hotspot, controlling several drought tolerance‐related traits has been introgressed into several popular cultivars using marker‐assisted backcrossing (MABC), and introgression lines giving significantly higher yield than the popular cultivars have been identified. Multiparent advanced generation intercross (MAGIC) approach has been found promising in enhancing genetic recombination and developing lines with enhanced tolerance to terminal drought and heat stresses.     

Genetic Advances in Adapting Rice to a Rapidly Changing Climate

Rice, with its wide geographic distribution extending from 50°N to 35°S, is expected to be the most vulnerable cultivated crop to future changing climates. Among the different abiotic stresses, extreme temperatures coinciding with critical developmental stages, increasingly frequent floods and drought spells, and worsening sea water inundation are some of the major threats to sustainable rice productivity. Following the successful implementation of molecular marker‐assisted backcrossing to introgress large‐effect QTL for submergence tolerance in rice mega varieties, rice breeding for drought, salinity and, recently, heat tolerance is employing the same approach. Although tolerance for combined submergence and salinity has been achieved, developing rice varieties with multiple tolerance for other abiotic and biotic stresses and finding the appropriate agronomic package to exploit their performance remain a challenge. The major bottleneck is the lack of unidentified large‐effect QTL for other abiotic stresses that are strongly influenced by genotype × environment (G × E) interaction. Rapid advances in the use of molecular tools, including a plethora of SNP markers, are expected to facilitate the development of major abiotic stress‐tolerant rice. In response to the actual farmer field situation, progress achieved in understanding and developing independent abiotic stress tolerance is being exploited to combine tolerances (for example, heat and drought; salinity and submergence) to address emerging environmental problems across a wide range of rice ecosystems.     

普通菜豆PvP5CS2基因对逆境胁迫的应答

为了探索逆境条件下脯氨酸积累的分子遗传机理, 改善作物的抗逆能力, 应用实时荧光定量PCR(RT-qPCR)技术和水杨酸法分别检测干旱、高盐(200 mmol L^-1 NaCl)和冷(4℃)胁迫条件下普通菜豆幼苗叶和根中脯氨酸合成酶基因PvP5CS2表达量和脯氨酸含量的变化。结果显示, 3种胁迫条件下普通菜豆幼苗叶和根中PvP5CS2的转录水平快速上升, 干旱处理4 d, 叶中PvP5CS2表达量达到最大值; 高盐处理下, 叶和根中PvP5CS2的表达高峰分别出现在2 h和6 h; 冷胁迫下, 叶和根中的表达高峰都出现在2 h; 随着胁迫时间的延长, PvP5CS2基因的转录水平逐渐下降。普通菜豆在逆境胁迫下, 幼苗叶和根中脯氨酸大量积累, 积累高峰出现在PvP5CS2基因表达高峰之后。这些结果说明, PvP5CS2基因的表达受干旱、高盐和冷胁迫诱导, 脯氨酸积累受PvP5CS2基因转录水平的调控。PvP5CS2基因在洋葱表皮细胞中的瞬时表达结果显示PvP5CS2蛋白定位在细胞核和膜上。     

银杏GbASR基因的克隆、生物信息学及表达分析

为了明确银杏ASR(ABA-stress-ripening)在应答非生物逆境胁迫中的作用,以便为ASR基因功能、环境胁迫研究提供材料。本研究以银杏(Ginkgo biloba)为材料,利用分子生物学技术对ASR基因进行了克隆,构建pCAMBIA1300-ASR-GFP融合表达载体,进行亚细胞定位分析。采用序列比对方法分析GbASR蛋白结构域及系统进化;利用qRT-PCR方法进行组织特异性表达及非生物逆境胁迫下的基因表达分析。结果表明,银杏ASR基因编码区全长546 bp,共编码182个氨基酸;蛋白质等电点为5.33,分子量大小为20111.24 Da,为亲水性稳定蛋白;GbASR蛋白含有1个高度保守的ABA/WDS结构域,且与火炬松PtASR蛋白同源性较高;GbASR蛋白定位于细胞核。此外,GbASR基因在银杏幼苗根、茎、叶各组织中均有表达,且在叶部有较高的表达;在高盐、干旱和高温胁迫下,GbASR基因均受诱导,且在不同胁迫处理时间0 h、6 h、24 h和48 h的表达存在显著的差异。以上分析说明GbASR基因参与响应高盐、干旱和高温等非生物胁迫,这为植物的育种及解释ASR基因功能提供依据。     

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。     

Genome-wide association analysis of seed germination percentage and germination index in <Emphasis Type="Italic">Brassica napus</Emphasis> L. under salt and drought stresses

Identifying genetic loci and possible candidate genes related to salt and drought stresses would be an economical, feasible and efficient way to accelerate the progress of abiotic tolerance breeding in rapeseed. In this study, a seed germination experiment using distilled water, salt and drought stresses was carried out with 520 B. napus germplasm resources. The rapeseed accessions showed large variation in the seed germination percentage (GP) and germination index (GI), with GP ranging from 26 to 100%, 0 to 100% and 0 to 100% and GI from 3.05 to 53.92, 0 to 24.50 and 0 to 25.00 in distilled water, under salt stress and drought stress, respectively. The coefficients of GP and GI were 0.72, 0.97 and 0.96 in distilled water, under salt stress and drought stress, respectively. A genome-wide association analysis of the 520 accessions was performed using the Q+K model and 31,839 single-nucleotide polymorphism (SNP) sites, revealing 23 SNPs associated with GP and 20 SNPs associated with GI under salt and drought stresses. The significant loci rs32406 and rs9466 on chromosome C08 and A04 were mapped to sites 3.32 and 2.20 kb from the salt- and drought-responsive gene BnaC08g41070 and drought-responsive gene BnaA04g02620D, respectively. BnaC06g01910D, a drought-responsive gene on chromosome C06, was found to overlap with the significant locus rs44427. In addition, 37 candidate genes associated with GP and GI under salt and drought stresses were detected. Most of these candidate genes regulate signal transduction and proline biosynthesis or encode a ubiquitin ligase E3; some have unknown biological functions and may respond to salt and drought stresses together as a complex network.     

水稻胆碱单加氧酶基因的克隆与表达分析

高等植物中甜菜碱由胆碱经两步氧化反应合成,其中由胆碱单加氧酶(CMO)催化第一步氧化反应,甜菜碱醛脱氢酶(BADH)催化第二步氧化反应.采用生物信息学技术和RT-PCR的方法从水稻幼苗组织中分离得到了水稻CMO基因的cDNA克隆,将其命名为OsCMO.该基因含有10个外显子和9个内含子,其开放阅读框编码一条长为410个氨基酸残基的多肽,与拟南芥、山菠菜、苋、碱蓬和甜菜等植物的胆碱单加氧酶的氨基酸序列的一致性50%～62%.mRNA分析表明OsCMO基因在水稻幼苗组织中的表达受高盐、低温和干旱等胁迫的上调诱导表达,表明该基因可能在抵御非生物胁迫中发挥重要作用.     

烟草细胞质小分子热激蛋白基因Nt- Hsp17.8的克隆及胁迫诱导表达特性分析

越来越多的研究表明,非生物胁迫影响植物的生长发育与栽培范围.为了通过转基因方法提高栽培番茄、马铃薯对热和干旱组合胁迫的抗性,利用RT-PCR结合RACE技术,从热处理的烟草叶片中分离出了细胞质小分子热激蛋白基因.分析表明,该基因cDNA全长876 bp,包含一个480 bp的开放阅读框,编码159个氨基酸,相对分子量约...     

Salicylic Acid and Abiotic Stress Responses in Rice

Among plant species rice (Oryza sativa L.) leaves can be characterised with a very high level of salicylic acid content; however, its exact role is still poorly understood. In the present work, rice genotypes with different levels of drought tolerance have been subjected to PEG‐induced drought or cold stress at 10 °C in order to find relationship between the salicylic acid metabolism and the level of stress tolerance; and between the salicylic acid level and other protective mechanisms. Although the drought‐sensitive genotypes usually contained slightly higher amount of salicylic acid than the tolerant ones, there was no strong correlation between the salicylic acid contents and the degree of drought tolerance. Because the expression pattern of the chorismate synthase and isochorismate synthase genes did not correlate with the level of salicylic acid, but there was a correlation between the levels of salicylic acid and ortho‐hydroxy‐cinnamic, it is assumed that the salicylic acid synthesis via ortho‐hydroxy‐cinnamic acid may play a more decisive role than the chorismate–isochorismate–salicylic acid pathway in rice. While the activity of the glutathione reductase enzyme did not show correlation with drought tolerance, the glutathione S‐transferase activities were usually higher in the leaves of the drought‐tolerant varieties than in the sensitive ones. The salicylic acid contents in the leaves were not substantially affected by the applied stress conditions; however, other stress‐related compounds polyamines showed marked, stress‐specific responses. Correlation data suggest that there is no direct link between the abiotic stress‐induced polyamine changes and the salicylic acid metabolism in rice.     

Expression analysis of BsAPase14 acid phosphatase,a stress responsive boiling-stable protein from <Emphasis Type="Italic">Triticum aestivum</Emphasis>

Developmental regulation and effect of abiotic stresses (osmotic, salt, heat, and drought) on induction of boiling-stable acid phosphatases (BsAPases) was studied in the growing wheat seedlings. SDS-PAGE revealed the induction of several boiling-stable polypeptides in a spatial and temporal manner in the growing seedlings. Western blot analysis, using polyclonal acid phosphatase antiserum, revealed the induction of a cross-reacting one 14 kDa boiling-stable acid phosphatase band (14 kDa, BsAPase14) in endosperm at 7 and 14 days of germination (DOG). Various abiotic stresses (heat, salt, and osmotic) also elicit accumulation of BsAPase14 in a tissue-dependent manner. Imposition of drought stress also resulted in an induction of boiling-stable protein (BsAPase-14) which disappeared after removing the stress. Based upon these observations, the possible role of BsAPase14 in water-stress tolerance is discussed.     

Acquired Thermo‐Tolerance and Trans‐Generational Heat Stress Response at Flowering in Rice

In rice, pre‐exposure to sublethal treatment followed by harsh lethal treatment is known to improve tolerance of different abiotic stresses at the vegetative stage within and across generations. Our major aim was to test the phenomenon of thermo‐tolerance at flowering across (trans)‐generations and within generation using rice cultivars contrasting for heat stress tolerance at flowering. To test trans‐generational response, plants were exposed to higher temperature at flowering stage and seeds obtained from previous generations were exposed to heat stress during flowering, which recorded significantly lower fertility when exposed to the same degree of stress in their subsequent generations. A pre‐acclimation to moderately high acclimating temperatures imposed over three different durations during the vegetative and initial reproductive stage showed positive response in the tolerant N22, particularly under severe heat stress (40 °C). This finding indicates the possibility of acquiring ameliorative thermo‐tolerant mechanisms at anthesis, restricted to tolerant genetic backgrounds to combat subsequent harsh conditions within the same generation. However, trans‐generational memory was ineffective in mitigating spikelet sterility losses in both tolerant and susceptible backgrounds. Rice is extremely sensitive to heat stress during flowering; hence, similar exercise across other crops of interest needs to be carried out before generalizing conclusions.     

Long‐Term Field Drought Affects Leaf Protein Pattern and Chloroplast Ultrastructure of Winter Wheat in a Cultivar‐Specific Manner

Recurrent drought periods of varying duration often cause extensive crop damage and affect wheat production in Southern Europe. This study compares biochemical and ultrastructural responses of four wheat (Triticum aestivum L.) cultivars to long‐term field drought, and their contribution to final grain yield. Gel electrophoresis and immunoblotting analyses combined with transmission electron microscopy and grain yield evaluation were employed to assess drought susceptibility of the wheat cultivars. Two of them behaved as drought‐tolerant, the other two presented as drought sensitive. Enhanced degradation of Rubisco large subunit (RLS), Rubisco small subunit (RSS) and Rubisco activase (RA) accompanied by an increased protease activity and reduced levels of heat shock proteins (HSP70) and dehydrins (DHNs) were associated with drought sensitivity. Drought tolerance coincided with relatively stable or increased HSP70 and DHN contents, and unchanged/higher levels of RLS, RSS and RA. Sensitive cultivars were more vulnerable to ultrastructural damages, showing obvious degradation of chloroplast membrane systems and depletion of leaf starch reserves. These drought responses affected yield potential, as tolerant cultivars gave higher yield under intense drought. Thus, our results provide additional insights into the complexity of plant drought responses, identifying multiple interacting traits that may serve as indirect selection criteria for wheat drought tolerance.     

Current status and future strategy in breeding chickpea for resistance to biotic and abiotic stresses

Chickpea (Cicer arietinum L.) production has remained static for the past two decades. One major limiting factor has been susceptibility of cultivars to several biotic and abiotic stresses that adversely affect yield. In recent years, cultivars resistant to Ascochyta blight (Ascochyta rabiei [Pass.] Lab.), Fusarium wilt (Fusarium oxysporum f. sp. ciceris), and cold have been bred and released in many countries. Some progress has been made in breeding for resistance to drought, insects, and cyst nematode, but not for viruses, heat, and salinity. Two or more stresses are of equal importance in most chickpea growing areas. Therefore, future efforts should be directed toward the development of cultivars with multiple-stress resistance. Proper understanding of important stresses in different countries and the genetics of resistance should lead to more systematic approaches to resistance breeding. Wild Cicer species hold promise and deserve attention in resistance breeding.     

玉米干旱诱导表达基因ZmCKS2的克隆与表达分析

在本实验室前期研究的干旱胁迫相关的抑制差减文库(SSH)中,发现一个玉米干旱胁迫诱导表达的EST序列,与拟南芥AtCKS2和AtCKS1基因有较高的同源性。应用生物信息学手段和同源克隆的方法,分离得到该基因,命名为ZmCKS2。序列分析表明该基因开放阅读框区267bp,编码88个氨基酸。ZmCKS2蛋白含有真核生物中高度保守的CKS(cyclin-dependent kinase regulatory subunit)结构域。应用在线分析软件预测该基因上游2kb序列表明,该序列具有启动子的核心序列和增强子序列,同时还具有与干旱等多种逆境胁迫相关的调控序列。应用实时荧光定量PCR分析表明该基因在幼穗中表达量最高,且受干旱和MeJA诱导上调表达,受低温和外源ABA诱导下调表达。