Effect of heat stress on wheat proteins during kernel development in wheat near-isogenic lines differing at Glu-D1

Two near-isogenic lines of the wheat variety Lance having Glu-D1a (HMW-GS 2 + 12) and Glu-D1d (HMW-GS 5 + 10) were subjected to several regimes of heat stress. In 2001, the temperature regimes were (i) 20/16 (day/night, °C) from planting to maturity, (ii) 20/16 except for a 3-day heat treatment of 35/20, 25 days after anthesis and (iii) 20/16 until 25 DAA, after which plants were subjected to 40/25 until maturity. In 2002, treatments (i) and (iii) were the same while treatment (ii) used a temperature of 40/25 °C for 3 days at 25 DAA. Seed was collected at 3-day intervals starting from 16 days after anthesis and analyzed for protein composition by SE-HPLC. The line with the Glu-D1d allele showed an earlier polymerization of glutenin than its allelic counterpart and a higher molecular weight of glutenin at maturity, this being deduced from measurements of the percentage of unextractable polymeric protein. It is postulated that the timing and rate of glutenin polymerization, and the timing of high temperature application may be the key factors contributing to an explanation of the effect of heat stress on functionality.     

大豆籽粒不同发育时期的转录组分析

为了从分子水平上研究大豆籽粒不同发育时期的油脂合成与累积机理,对大豆开花20 d(DD20)、30 d(DD30)、40 d(DD40)、50 d(DD50)的籽粒进行了转录组测序。通过对转录组测序数据的分析,共得到原始数据461 566 988条,经过滤获得开花后20,30,40和50 d的clean reads,分别为107 548 920,111 670 776,109 339 672和108 884 270条。DD20/DD30、DD30/DD40、DD40/DD50比较组中的差异表达基因分别为4 759,6 245和13 763个,其中上调基因分别为1 801,2 941和5 695个。差异表达基因的KEGG pathway分析中,分别得到134,133和136条代谢通路,筛选到8个与油脂合成相关的差异表达基因,ACC、FATB、GPAT、DGAT1、G3PDH、KASI、SAD和FAD2。研究结果对深入研究大豆籽粒脂质合成的调控机理及大豆高油育种具有重要参考价值。     

Transcriptome analysis of salt-stress response in three seedling tissues of common wheat

Xiaoyan 60(XY60) is a new wheat variety bred in the laboratory of Zhensheng Li.After salt treatment, seedlings of XY60 maintain green leaves and produce longer roots than the high yielding cultivar Zhongmai 175(ZM175).To explain these different phenotypes we carried out an RNA-Seq analysis using 12 samples from three tissues of both varieties subjected to salt and control treatments.By comparing data from the salt treated plants with the control, 703, 979, and 1197 differentially expressed genes(DEGs) were detected in new leaves, old leaves, and roots of XY60, respectively.The corresponding numbers for ZM175 were 613, 1401, and 1301.The most significantly enriched Gene Ontology(GO) terms and KEGG pathways were associated with polyunsaturated fatty acid(PUFA) metabolism in both new and old leaves from XY60.They were associated with photosynthesis and energy metabolism in ZM175.The most significantly enriched KEGG pathway in roots of both varieties was “glucosinolate biosynthesis”.In addition, jasmonic acid(JA) concentration in XY60 was higher than in ZM175, although it increased significantly in both varieties following salt treatment.Trends in relative expression levels of AOS, MYC2, and JAZ revealed by qRT-PCR were concordant with those from RNA-Seq.Our results suggest that PUFAs may contribute to salt tolerance in common wheat by enhancing the photosynthetic system and JA-related pathways.     

Malondialdehyde content as a potential biochemical indicator of tolerant Cenchrus ciliaris L. genotypes under heat stress treatment

Buffel grass (Cenchrus ciliaris L.) is an important forage grass in arid and semiarid regions. As part of a genetic improvement programme, four genotypes [Biloela (Bl), Americana (Am), Texas (Tx) and Sexual (Sx)] were categorized by tolerance to heat stress in a greenhouse experiment. At 30 d after sowing, half of the plants (control plants) were transferred to growth chambers (28°C day per night), and the other half (pre‐treated plants, Prt) were exposed to heat stress treatment (0, 24, 48 and 72 h; 45°C day per night). Malondialdehyde (MDA) content, an indicator of oxidative damage, was determined from foliar samples. During heat stress, Sx showed the earliest increase in MDA (at 24 h) followed by Tx (48 h) and Am and Bl (72 h). Results were compared with heat‐stress tolerance evaluated as morphological traits at the end of recuperation (60 d after sowing). Fresh weight and aerial plant height were lowest in the Prt‐Sx genotype and highest in Am and Bl genotypes; Tx showed intermediate tolerance. Results suggest that tolerance to heat stress in C. ciliaris genotypes could be related to the capacity for regulating the oxidative damage increase. Foliar MDA content might therefore be used in a genetic improvement programme of C. ciliaris as a potential biochemical indicator for a rapid, simple and low‐cost identification of heat‐stress tolerant genotypes.     

不同基因型油菜对苗期水分胁迫的生理响应

通过盆栽试验分析了17个不同基因型甘蓝型油菜对苗期水分胁迫的生理响应及抗旱性差异。结果表明,水分胁迫下不同油菜材料9个生理生化指标的抗旱系数（Dc）存在显著差异,其中叶片萎蔫指数（WI）的变异系数和改良潜力最大。17份材料的平均隶属函数值（AS）变幅在0.12～0.94之间。相关分析表明AS与9个生理生化指标达显著或极显著相关,其中相关性最高的是WI和叶绿素含量,而且WI还与其余的8个生理生化指标具有很高的相关性。利用隶属函数法和系统聚类法对供试材料的抗旱性进行了评价分级,发现两种分类方法的评价结果一致,均可将供试油菜材料划分为不抗、低抗、中抗和高抗4种类型。鉴定获得了华杂10号、B108、Q2和中双7号等4份高度抗旱材料（AS > 0.75）。由于WI与苗期综合抗旱性高度相关,且测定简便,因此叶片萎蔫指数可作为油菜苗期抗旱性鉴定的关键指标。     

大麦根系响应湿害胁迫的转录组分析

湿害是影响大麦产量和品质的主要非生物胁迫之一。为进一步明确大麦响应湿害胁迫的应答机理,本研究以耐湿大麦品种泰兴9425为材料,利用高通量转录组测序(RNA-seq)技术,比较不同湿害胁迫时间下的基因表达差异。结果表明,湿害胁迫12 h后获得1 436个差异表达基因,而胁迫48 h后获得2 090个差异表达基因,其中共同上调表达基因286个。GO富集分析发现,涉及代谢过程、细胞膜、催化活性等的差异表达基因占比最多。KEGG富集分析发现,差异表达基因主要富集于糖酵解、类黄酮生物合成、乙烯合成等代谢通路。对7个耐湿相关的候选基因进行qRT-PCR分析,发现差异表达基因的表达量变化趋势与RNA-seq结果一致。     

营养生长期持续高温处理对玉米叶片转录组及生化指标的影响

以玉米杂交种郑单958和登海605为材料,通过转录组测序分析玉米营养生长期叶片受热胁迫后基因差异表达、基因功能富集和KEGG通路富集,使用微量法和HPLC法分析叶片中相关生化指标的变化。结果表明,高温胁迫会降低光合作用关键酶二磷酸核酮糖羧化酶(Rubisco)和三羧酸循环限速酶柠檬酸合酶(CS)的活力,升高丙二醛(MDA)、脱落酸(ABA)和赤霉素(GA₃)的含量,降低水杨酸(SA)的含量。与常温对照比,玉米营养生长期叶片受热胁迫后,两个品种4个比较组合累计检测到16 996个差异表达的基因,共同差异表达的基因有1 160个。光合作用、细胞代谢、叶绿体膜和质体膜等相关基因的表达会响应热胁迫。     

The impact of emmer genetic diversity on grain protein content and test weight of hexaploid wheat under high temperature stress

High temperature has a negative impact on wheat grain quality and reduces market value. Emmer wheat (Triticum dicoccon Schrank), one of the earliest domesticated wheat species, is a source of genetic diversity for the improvement of heat and drought tolerance in modern wheat. However, the potential of emmer wheat for the improvement of grain physical quality under high temperature stress is little studied. A diverse set of 184 emmer-based hexaploid lines was developed by crossing emmer wheat with hexaploid wheat and backcrossing once to hexaploid wheat. These materials, seven hexaploid recurrent parents and seven commercial cultivars, were evaluated at two times of sowing (E1 and E2) in the field, in 2015–2016. The materials were genotyped using a 90 K SNP platform and these data were used to estimate the contribution of emmer wheat to the progeny. Significant phenotypic and genetic variation for grain physical quality traits including protein content and test weight was observed. High temperature significantly increased protein content and decreased test weight. Large scale field phenotyping identified emmer progenies with improved grain characteristic compared to their respective parents and commercial cultivars in both environments. A few families consistently produced higher trait means across environments compared to their recurrent parents. The emmer wheat parent contributed between 1 and 37% of the genome in emmer-based genotypes. Selected emmer derived lines with superior protein content and test weight, tended to have a greater genetic contribution from the emmer parent, ranging from 12 to 37% and 7–37% in E1 and E2, respectively. It was concluded that new genetic variation for seed traits, such as protein content and test weight, can be introduced to hexaploid wheat from emmer wheat. The newly developed emmer derivatives identified with enhanced grain quality under high temperature stress can potentially be used to improve grain quality through breeding.     

花后高温胁迫下匀播春小麦旗叶的转录组分析

为研究不同播种模式下花后高温胁迫对春小麦旗叶转录组的影响,以宁春50号为试验材料,采用人工模拟高温的方法,设条播和匀播两种播种方式,灌水施肥方式均为水肥一体化,对高温处理后的春小麦旗叶分别构建转录组测序文库,采用FPKM法计算基因的相对表达量,并对差异表达基因进行KEGG通路分析。结果表明,相较于常温处理,高温胁迫下条播和匀播滴灌处理分别有199和1 819个基因上调表达,55和1 335个基因下调表达,说明高温胁迫下匀播滴灌处理对春小麦旗叶转录组的表达影响明显。KEGG通路富集分析结果显示,高温胁迫下与条播滴灌处理对比,匀播滴灌处理的春小麦光合作用-天线蛋白通路注释到的差异表达基因最多,有52个,其次为淀粉和蔗糖通路(注释到50个差异表达基因);而常温条件下与条播滴灌处理对比,匀播滴灌处理的春小麦淀粉和蔗糖代谢通路注释到的差异表达基因最多,有49个,其次为光合作用-天线蛋白通路(注释到41个差异表达基因)。因此,高温胁迫下匀播滴灌技术可以更好地改善植物的光合系统,有效缓解高温引起的小麦植株早衰。     

Gibberellin-sensitive Rht alleles confer tolerance to heat and drought stresses in wheat at booting stage

The present study compared six near-isogenic lines from different backgrounds, varying in Rht (reduced height) alleles for gibberellin (GA) response, for heat and drought tolerance. Various temperature (day/night: 20/12 °C to 39/31 °C), and irrigation (100% field capacity and no irrigation) treatments were imposed at booting stage. Results depicted a 50% reduction in grain yield at 36.7 °C with irrigation and at 31.2 °C without irrigation. Number of grains per spike was severely reduced over 26 °C for water-stressed plants while irrigated plants did not show any significant reduction up to 34 °C. Ear numbers per pot and spikelets per spike were unaffected by irrigation till 33 °C. In Mercia, grain nitrogen, sulphur, and Hagberg falling number (HFN) were significantly higher for severe dwarfs (Rht-D1c and Rht-12), however, in Maris Widgeon, rht (tall) recorded higher nitrogen and sulphur concentration while Rht-B1b presented the maximum sedimentation of sodium dodecyl sulphate (SDS). The SDS was higher in Maris Widgeon while HFN were superior in Mercia. Conclusively, the present study demonstrated that heat and drought stresses severely reduced the grain yield while improved grain quality traits. The GA-insensitive semi dwarf Rht-B1b was the most promising genotype for grain yield and HFN under combined drought and heat stresses.     

Response of chickpea genotypes to low temperature stress during reproductive development

Low temperature at flowering is a major constraint to improved yield of chickpea in many regions of the world. In particular, cool dryland environments such as southern Australia, parts of the Indian sub-continent and the Mediterranean would benefit from cultivars with the ability to flower and set pods early in the growing season before soil moisture becomes a limiting factor. This paper demonstrates that low temperature (less than 15 °C) affects both the development and function of reproductive structures in the chickpea flower. Aspects of the male and the female gametophyte phases of development are described. Comparisons between chilling sensitive genotypes are made to identify the likely causes of flower abortion. The function of pollen derived from chilling sensitive plants is clearly affected most by low temperature stress, particularly the growth of the pollen tubes down the style before fertilisation occurs. In contrast, pollen tubes derived from chilling tolerant plants continue to grow down the style under low temperature stress. Although other stages of development and function were affected by low temperature, including sporogenesis, pollen germination, and the stigma, none were correlated to the phenotype of the mother plant. The implications of the findings for chickpea improvement programmes are discussed.     

利用转录组测序对不同贮藏温度处理玉米种子的苗期差异表达基因分析

利用常温与低温贮藏3个月的玉米杂交种种子进行标准发芽试验,取发芽7d玉米幼苗进行转录组测序分析,通过GO、KOG、KEGG数据库分析处理差异表达基因的功能和参与的调控路径.以常温贮藏处理为参考,对转录组测序数据进行比较,低温贮藏处理共获得35个差异表达基因,其中上调基因28个、下调基因7个.在GO功能注释分析中,有26...     

Identification of CIMMYT spring bread wheat germplasm maintaining superior grain yield and quality under heat-stress

Unpredictable temperatures and rainfall associated with climate change are expected to affect wheat (Triticum aestivum L.) production in various countries. The development of climate-resilient spring wheat cultivars able to maintain grain yield and quality is essential to food security and economic returns. We tested 54 CIMMYT spring bread wheat genotypes, developed and/or released over a span of 50 years, in the field for two years under optimum sowing dates, as well as using two delayed sowing dates to expose crops to medium and severe heat-stress conditions. The grain yield and yield components were severely affected as the heat-stress increased. Two contrasting groups of 10 lines each were identified to determine the effect of heat-stress on bread-making quality. The first set included entries that produced high yields in optimal conditions and maintained higher yields under heat-stress (superior-yielding lines), while the second set included genotypes that did not perform well in the environment with high temperature (inferior-yielding lines). We identified genotypes exhibiting bread-making quality stability, as well as the quality traits that had higher correlation with the loaf volume in the environment without stress and under heat-stress. Of all the quality traits tested, dough extensibility (AlvL) and grain protein content had a significant influence in heat-stress adaptation. Most of the lines from the superior-yielding group were also able to maintain and even improve quality characteristics under heat-stress and therefore, could be used as parents in breeding to develop high-yielding and stable quality wheat varieties.     

Heat and drought stress on durum wheat: Responses of genotypes,yield, and quality parameters

Heat and/or drought stress during cultivation are likely to affect the processing quality of durum wheat (Triticum turgidum L. ssp. durum). This work examined the effects of drought and heat stress conditions on grain yield and quality parameters of nine durum wheat varieties, grown during two years (2008–09 and 2009–10). Generally, G and E showed main effects on all the parameters whereas the effects of G × E were relatively small. More precipitation in Y09–10 may account for the large differences in parameters observed between crop cycles (Y08–09 and Y09–10). Combined results of the two crop cycles showed that flour protein content (FP) and SDS sedimentation volume (SDSS) increased under both stress conditions, but not significantly. In contrast the gluten strength-related parameters lactic acid retention capacity (LARC) and mixograph peak time (MPT) increased and decreased significantly under drought and heat stress, respectively. Drought and heat stress drastically reduced grain yield (Y) but significantly enhanced flour yellowness (FY). LARC and the swelling index of glutenin (SIG) could be alternative tests to screen for gluten strength. Genotypes and qualtiy parameters performed differently to drought and heat stress, which justifies screening durum wheat for both yield and quality traits under these two abiotic stress conditions.     

Winter wheat genotypes under different levels of nitrogen and water stress: Changes in grain protein composition

Hard white winter wheat with superior and consistent quality is preferable for Asian markets. This study investigated the combined influences of moisture deficit during grain-fill and N management on protein quality, dough rheological properties, and protein molecular weight distributions in soft and hard winter wheats. Genotypes were grown under an irrigation gradient and two N-fertilization levels. Grain polyphenol oxidase (PPO) activity, SDS sedimentation, and Mixograph analyses were evaluated. Flour protein composition was characterized using SE-HPLC. Moisture stress during grain-fill increased flour protein content. N fertilization increased flour protein content. No significant correlation was found between flour protein and PPO. Changes in protein composition were related to general increases in protein content, regardless if the result of reduced irrigation or increased fertilization rate. The percentage of monomeric proteins increased more than the polymeric proteins as flour protein increased. Similarly, SDS sedimentation volume increased as a function of protein content. As expected, subunit GluD1 5+10 was associated with larger sedimentation volume and higher dough strength in genotypes as compared to those with subunit GluD1 2+12. Biplot analyses showed that genotypes of similar protein quality and composition responded similarly to N and irrigation treatments.     

低磷胁迫下玉米幼苗生理响应及相关基因表达研究

以玉米品种合344为试验材料,对低磷胁迫下玉米幼苗叶片生理表型指标与玉米叶片转录组差异基因表达数据联合分析,探究低磷胁迫下与玉米幼苗叶片生理表型变化相关的关键响应基因及其调控规律.结果表明,低磷胁迫下玉米叶片的叶绿素荧光参数(FV/FM)以及叶绿素含量均有不同程度的降低,并且叶绿素合成相关基因大部分为下调表达,叶绿素降...     

干旱胁迫下过表达TaER小麦的转录组及光合特性分析

为探究小麦TaER基因调控蒸腾效率及抗旱性的分子机制,以2个T_3代转TaER基因株系及其野生型为材料,对其正常灌溉和干旱胁迫处理下的灌浆期旗叶进行转录组(RNA-seq)测序,分析其差异表达基因并进行GO和KEGG富集分析,探究其功能及可能参与的调控通路,同时测定光合及蒸腾参数。结果表明,与野生型相比,TaER过表达株系在正常灌溉和干旱胁迫下分别获得1 439和607个共同的差异表达基因;GO富集分析发现,光合作用、脂质和膜脂代谢基因响应干旱胁迫;KEGG富集分析表明,亚油酸和α-亚麻酸代谢中的酯氧合酶、甘油脂代谢中的糖基转移酶基因均上调表达,光合作用相关基因响应干旱胁迫;干旱胁迫下,TaER过表达株系的净光合速率和水分利用效率较高,蒸腾速率较低。综上所述,TaER过表达株系可能是通过调控脂类代谢及光合作用等生物过程,提高光合速率及水分利用效率,从而适应干旱胁迫的。