一氧化氮调节盐胁迫下小麦幼苗根部质膜H+-ATPase和焦磷酸酶活性提高耐盐性

采用外源一氧化氮(NO)供体硝普钠(SNP)研究了NO对盐胁迫下小麦(Triticum aestivum L.)幼苗耐盐性的影响.结果表明,0.1 mmol/L SNP处理显著缓解了1 50 mmol/L NaCl胁迫对小麦幼苗生长的抑制效应,包括水分丧失以及叶绿素降解,从而提高了小麦幼苗的耐盐性.进一步结合1 mg/mL血红蛋白处理则显著逆转了SNP诱导的上述效应;利用亚硝酸钠和铁氰化钾作为对照也证实了NO对小麦幼苗耐盐性的专一性调节作用,并可能与NO对小麦幼苗根部质膜H -ATPase和焦磷酸酶活性诱导有关.此外,尽管NO显著提高了盐胁迫下小麦幼苗根部细胞质膜H -ATPase和焦磷酸酶的ATP水解活性,但是对跨膜H 转运则没有明显影响.应用外源CaSO4和EGTA处理也证实,Ca2 可能在NO诱导的质膜H -ATPase和焦磷酸酶活性的提高过程中起信号作用.另外,分析盐胁迫下小麦幼苗根部Na 和K 含量的变化也发现,NO对Na 含量没有明显影响,但是却显著提高了K 水平和K /Na 比,这可能也是NO提高小麦幼苗耐盐性的原因之一.     

一氧化氮调节盐胁迫下小麦幼苗根部质膜H^＋-ATPase和焦磷酸酶活性提高耐盐性

采用外源一氧化氮(NO)供体硝普钠(SNP)研究了NO对盐胁迫下小麦(Triticum aestivum L.)幼苗耐盐性的影响。结果表明,0.1 mmol/L SNP处理显著缓解了150 mmol/L NaCl 胁迫对小麦幼苗生长的抑制效应,包括水分丧失以及叶绿素降解,从而提高了小麦幼苗的耐盐性。进一步结合1 mg/mL血红蛋白处理则显著逆转了SNP诱导的上述效应;利用亚硝酸钠和铁氰化钾作为对照也证实了NO对小麦幼苗耐盐性的专一性调节作用,并可能与NO对小麦幼苗根部质膜 H -ATPase和焦磷酸酶活性诱导有关。此外,尽管NO显著提高了盐胁迫下小麦幼苗根部细胞质膜H -ATPase和焦磷酸酶的ATP水解活性,但是对跨膜H 转运则没有明显影响。应用外源CaSO4 和 EGTA 处理也证实,Ca2 可能在NO诱导的质膜 H -ATPase和焦磷酸酶活性的提高过程中起信号作用。另外,分析盐胁迫下小麦幼苗根部 Na 和K 含量的变化也发现,NO对Na 含量没有明显影响,但是却显著提高了K 水平和K /Na 比,这可能也是NO提高小麦幼苗耐盐性的原因之一。     

不同耐盐性小麦根Na~+和K~+的吸收特性

以耐盐小麦品种‘德抗961’和盐敏感小麦品种‘鲁麦15’为材料,研究小麦根Na+、K+吸收特性及其与耐盐性关系。结果表明,2个小麦品种根K+吸收动力学曲线均符合Michaelis-Menten方程,即V=Vmax×[S]/([S]+Km)+k×[S]。低浓度(低于25mmol·L-1)NaCl处理对根高亲和K+吸收系统转运K+具有促进作用,对耐盐品种‘德抗961’的促进作用更大。小麦根高亲和K+吸收系统是通过K+/H+同向转运,而不是K+/Na+同向转运。NaCl处理对根低亲和K+吸收系统有抑制作用,对盐敏感品种‘鲁麦15’的抑制作用更大。NaCl处理导致2个小麦品种根和叶片中的K+含量显著下降,Na+含量显著升高,但‘德抗961’根和叶片中的K+含量均显著高于‘鲁麦15’,‘德抗961’根中Na+含量显著高于‘鲁麦15’,而其叶片中Na+含量显著低于‘鲁麦15’,从而保证NaCl胁迫下其叶片较高的K+/Na+比。非选择性阳离子通道是小麦根Na+吸收的主要途径,K+通道是Na+吸收的一条重要途径。这些结果表明小麦部分通过调节根系K+吸收系统而维持叶片较高的K+/Na+比,从而提高其耐盐性。     

人工海水胁迫下小麦种质资源的耐盐性筛选与鉴定

利用人工配制的海水筛选耐盐性较好的小麦品种,为沿海滩涂地区的小麦耐盐育种提供重要信息。本研究利用人工海水处理的方法,对363份小麦种质资源进行了芽期耐盐性初步鉴定,筛选出芽期耐盐性为1级的小麦种质28份。进一步对芽期耐盐性较好的48份小麦种质进行了苗期耐盐性鉴定,并对其耐盐指标进行隶属值模糊评价分析,从中鉴定出了2个苗期耐盐性较强的小麦种质,分别为淮麦31和红壳洋麦。依据来源的不同,发现小麦种质资源的芽期耐盐性大小依次为地方品种>育成品种>国外引进品种。小麦芽期与苗期的耐盐性相关分析表明,二者相关性极低(r=-0.0051)。     

小麦耐低磷相关性状的全基因组关联分析

通过对小麦耐低磷相关性状进行全基因组关联分析(GWAS,genome-wide association study),挖掘与小麦耐低磷性显著相关的单核苷酸多态性标记(SNP,single nucleotide polymorphism)位点及候选基因,为小麦耐低磷性状的遗传基础和分子机制研究提供理论参考。本试验以198份黄淮麦区小麦品种(系)为试验材料,设置低磷和正常磷营养液水培试验,利用小麦35K芯片对分布于小麦全基因组的11896个SNP,采用Q+K关联模型对小麦耐低磷性相关性状进行关联分析。结果表明,小麦耐低磷性状表现出广泛的表型变异,变异系数为15.65%~26.59%,多态性信息含量(PIC,polymorphic information content)为0.095~0.500。群体结构分析表明,试验所用自然群体可分为2个亚群,GWAS共检测到67个与小麦耐低磷相关性状显著关联的SNP位点(P≤0.001),这些位点分布在除3A、3B和3D以外的18条染色体上,单个SNP位点可解释5.826%~9.552%的表型变异。在这些显著位点中有4个SNP位点同时关联到了2个不同的耐低磷性状。对67个SNP位点进行发掘,筛选到7个可能与小麦耐低磷性有关的候选基因。TraesCS6A02G001000和TraesCS6A02G001100在锌指合成中有重要作用;TraesCS6A02G118100可能为低磷胁迫诱导基因;TraesCS5D02G536400、TraesCS1B02G154200和TraesCS5D02G536500与低磷胁迫相关酶类基因家族有关;TraesCS1D02G231200与植物DUF 538结构域蛋白有关,是植物胁迫相关调控蛋白候选基因。     

一氧化氮对盐胁迫下小麦幼苗根生长和氧化损伤的影响

0.05和0.10 mmol/L一氧化氮(NO)供体硝普钠(sodium mtropmsside,SNP)处理明显减轻NaCl浓度为150 mmo1/L左右的盐胁迫对小麦幼苗根生长的抑制效应,其中0.05mmol/L的SNP效果最明显;0.30mmol/L以上的SNP处理对根抑制无明显缓解作用;当NaCl浓度大于300 mmol/L时,各种浓度的SNP均不能减轻盐胁迫对根生长的抑制.以N O清除剂血红蛋白(hemoglobin,Hb)以及NOx-,K3Fe(CN)6等为对照,观察到0.05 mmol/L的SNP能不同程度地提高150mmo/L盐胁迫下小麦幼苗根尖细胞中超氧化物歧化酶(SOD)、过氧化物酶(POD)和抗坏血酸过氧化物酶(ascorbateperoxidase,APX)活性,明显降低MDA、H2O2和O2-.的积累,阻断盐胁迫诱导的根尖细胞DNA片段化,表明NO能有效缓解盐胁迫引起的小麦幼苗根尖细胞的氧化损伤.     

盐胁迫下小麦甜菜碱和脯氨酸含量变化

运用高效液相色谱-质谱(HPLC-MS)联用技术分析了耐盐性强、中、弱3个小麦品种SW12、宁春4号和中国春苗期5个NaCl浓度胁迫下甜菜碱和脯氨酸含量的变化.方差分析表明盐胁迫下3个小麦品种之间甜菜碱的含量差异达到极显著水平(P＜0.01),SW12的含量最高,宁春4号次之,中国春最低,与小麦耐盐性表现相一致;脯氨酸在叶片中的含量差异不显著,在根中宁春4号和中国春的含量有显著差异(P＜0.05).结果表明:小麦叶和根中甘氨酸甜菜碱含量与小麦盐胁迫呈正相关,是小麦体内抵御盐胁迫的渗透调节物质之一,可作为小麦耐盐性鉴定指标.     

一种以幼根K+外漏量鉴定小麦耐盐性的方法

经100 mmol·L-1 NaCl处理的小麦幼根、胚芽和胚芽鞘的K+净外漏量和K+净外漏率显著增加,盐敏感小麦品种'LM15'的幼根K+净外漏量和净外漏率明显大于耐盐品种'DK961';2个品种小麦幼根K+外漏率差异主要是由其成熟区引起的.幼根K+净外漏量及净外漏率均与小麦的耐盐性呈负相关,因此认为幼根K+外漏可以作为快速鉴定NaCl胁迫下小麦耐盐性的指标.     

一氧化氮供体硝普钠浸种缓解盐胁迫对小麦种子萌发的抑制作用

以小麦品种‘德抗961＇为材料,用NO供体硝普钠（SNP）浸种研究外源NO对盐胁迫下小麦种子萌发的影响.结果表明：0.06 mmol/L的SNP浸种24 h后对盐胁迫下小麦种子发芽率、发芽指数、活力指数和吸胀速率的下调都有显著缓解作用;SNP浸种对盐胁迫下α-淀粉酶的活性无明显影响,但能显著提高盐胁迫下β-淀粉酶的活性;进一步研究表明,SNP浸种预处理对盐胁迫下的α-淀粉酶同工酶变浅的条带有所恢复（尤其是条带3）,同时使盐胁迫下变浅的β-淀粉酶同工酶的条带有明显的恢复（尤其是d、e、f、g）.并且SNP能显著降低盐胁迫下小麦地上部分和根中的Na^＋含量,提高其K＋含量,从而使K^＋/Na^＋显著提高.以上结果表明：SNP浸种预处理提高盐胁迫下小麦种子的萌发,主要是通过提高β-淀粉酶的活性来实现的.     

基于叶片生理指标的小麦芽期耐盐性评价

土壤盐渍化严重影响小麦生产,提高小麦耐盐性是应对土壤盐渍化的主要生物途径之一.小麦芽期亦是对盐分较为敏感的时期,小麦芽期耐盐性的强弱对盐碱地小麦种植至关重要.为探讨利用叶片生理指标进行小麦芽期耐盐性评价的可行性,该文以沧麦6005及其73个叠氮化钠诱变家系为试验材料,在超纯水和40％人工海水条件下,对芽期叶片中脯氨酸、...     

A large‐scale genomic association analysis identifies the candidate causal genes conferring stripe rust resistance under multiple field environments

The incorporation of resistance genes into wheat commercial varieties is the ideal strategy to combat stripe or yellow rust (YR). In a search for novel resistance genes, we performed a large‐scale genomic association analysis with high‐density 660K single nucleotide polymorphism (SNP) arrays to determine the genetic components of YR resistance in 411 spring wheat lines. Following quality control, 371 972 SNPs were screened, covering over 50% of the high‐confidence annotated gene space. Nineteen stable genomic regions harbouring 292 significant SNPs were associated with adult‐plant YR resistance across nine environments. Of these, 14 SNPs were localized in the proximity of known loci widely used in breeding. Obvious candidate SNP variants were identified in certain confidence intervals, such as the cloned gene Yr18 and the major locus on chromosome 2BL, despite a large extent of linkage disequilibrium. The number of causal SNP variants was refined using an independent validation panel and consideration of the estimated functional importance of each nucleotide polymorphism. Interestingly, four natural polymorphisms causing amino acid changes in the gene TraesCS2B01G513100 that encodes a serine/threonine protein kinase (STPK) were significantly involved in YR responses. Gene expression and mutation analysis confirmed that STPK played an important role in YR resistance. PCR markers were developed to identify the favourable TraesCS2B01G513100 haplotype for marker‐assisted breeding. These results demonstrate that high‐resolution SNP‐based GWAS enables the rapid identification of putative resistance genes and can be used to improve the efficiency of marker‐assisted selection in wheat disease resistance breeding.     

Genome-wide association mapping of salinity tolerance in rice (Oryza sativa)

Salinity tolerance in rice is highly desirable to sustain production in areas rendered saline due to various reasons. It is a complex quantitative trait having different components, which can be dissected effectively by genome-wide association study (GWAS). Here, we implemented GWAS to identify loci controlling salinity tolerance in rice. A custom-designed array based on 6,000 single nucleotide polymorphisms (SNPs) in as many stress-responsive genes, distributed at an average physical interval of <100 kb on 12 rice chromosomes, was used to genotype 220 rice accessions using Infinium high-throughput assay. Genetic association was analysed with 12 different traits recorded on these accessions under field conditions at reproductive stage. We identified 20 SNPs (loci) significantly associated with Na⁺/K⁺ ratio, and 44 SNPs with other traits observed under stress condition. The loci identified for various salinity indices through GWAS explained 5–18% of the phenotypic variance. The region harbouring Saltol, a major quantitative trait loci (QTLs) on chromosome 1 in rice, which is known to control salinity tolerance at seedling stage, was detected as a major association with Na⁺/K⁺ ratio measured at reproductive stage in our study. In addition to Saltol, we also found GWAS peaks representing new QTLs on chromosomes 4, 6 and 7. The current association mapping panel contained mostly indica accessions that can serve as source of novel salt tolerance genes and alleles. The gene-based SNP array used in this study was found cost-effective and efficient in unveiling genomic regions/candidate genes regulating salinity stress tolerance in rice.     

A genome-wide association study uncovers novel genomic regions and candidate genes of yield-related traits in upland cotton

Key message

A total of 62 SNPs associated with yield-related traits were identified by a GWAS. Based on significant SNPs, two candidate genes pleiotropically increase lint yield.

Abstract

Improved fibre yield is considered a constant goal of upland cotton (Gossypium hirsutum) breeding worldwide, but the understanding of the genetic basis controlling yield-related traits remains limited. To better decipher the molecular mechanism underlying these traits, we conducted a genome-wide association study to determine candidate loci associated with six yield-related traits in a population of 719 upland cotton germplasm accessions; to accomplish this, we used 10,511 single-nucleotide polymorphisms (SNPs) genotyped by an Illumina CottonSNP63K array. Six traits, including the boll number, boll weight, lint percentage, fruit branch number, seed index and lint index, were assessed in multiple environments; large variation in all phenotypes was detected across accessions. We identified 62 SNP loci that were significantly associated with different traits on chromosomes A07, D03, D05, D09, D10 and D12. A total of 689 candidate genes were screened, and 27 of them contained at least one significant SNP. Furthermore, two genes (Gh_D03G1064 and Gh_D12G2354) that pleiotropically increase lint yield were identified. These identified SNPs and candidate genes provide important insights into the genetic control underlying high yields in G. hirsutum, ultimately facilitating breeding programmes of high-yielding cotton.

     

Genome-wide association mapping reveals key genomic regions for physiological and yield-related traits under salinity stress in wheat (Triticum aestivum L.)

A genome-wide association study (GWAS) was conducted using six different multi-locus GWAS models and 35K SNP array to demarcate genomic regions underlying reproductive stage salinity tolerance. Marker-trait association analysis was performed for salt tolerance indices (STI) of 11 morpho-physiological traits, and the actual concentrations of Na⁺ and K⁺, and the Na⁺/K⁺ ratio in flag leaf. A total of 293 significantly associated quantitative trait nucleotides (QTNs) for 14 morpho-physiological traits were identified. Of these 293 QTNs, 12 major QTNs with R² ≥ 10.0% were detected in three or more GWAS models. Novel major QTNs were identified for plant height, number of effective tillers, biomass, grain yield, thousand grain weight, Na⁺ and K⁺ content, and the Na⁺/K⁺ ratio in flag leaf. Moreover, 48 candidate genes were identified from the associated genomic regions. The QTNs identified in this study could potentially be targeted for improving salinity tolerance in wheat.     

Control of sodium transport in durum wheat

In many species, salt sensitivity is associated with the accumulation of sodium (Na(+)) in photosynthetic tissues. Na(+) uptake to leaves involves a series of transport steps and so far very few candidate genes have been implicated in the control of these processes. In this study, Na(+) transport was compared in two varieties of durum wheat (Triticum turgidum) L. subsp. durum known to differ in salt tolerance and Na(+) accumulation; the relatively salt tolerant landrace line 149 and the salt sensitive cultivar Tamaroi. Genetic studies indicated that these genotypes differed at two major loci controlling leaf blade Na(+) accumulation (R. Munns, G.J. Rebetzke, S. Husain, R.A. James, R.A. Hare [2003] Aust J Agric Res 54: 627-635). The physiological traits determined by these genetic differences were investigated using measurements of unidirectional (22)Na(+) transport and net Na(+) accumulation. The major differences in Na(+) transport between the genotypes were (1) the rate of transfer from the root to the shoot (xylem loading), which was much lower in the salt tolerant genotype, and (2) the capacity of the leaf sheath to extract and sequester Na(+) as it entered the leaf. The genotypes did not differ significantly in unidirectional root uptake of Na(+) and there was no evidence for recirculation of Na(+) from shoots to roots. It is likely that xylem loading and leaf sheath sequestration are separate genetic traits that interact to control leaf blade Na(+).     

Genome-Wide Association Study for Wool Production Traits in a Chinese Merino Sheep Population

Genome-wide association studies (GWAS) provide a powerful approach for identifying quantitative trait loci without prior knowledge of location or function. To identify loci associated with wool production traits, we performed a genome-wide association study on a total of 765 Chinese Merino sheep (JunKen type) genotyped with 50 K single nucleotide polymorphisms (SNPs). In the present study, five wool production traits were examined: fiber diameter, fiber diameter coefficient of variation, fineness dispersion, staple length and crimp. We detected 28 genome-wide significant SNPs for fiber diameter, fiber diameter coefficient of variation, fineness dispersion, and crimp trait in the Chinese Merino sheep. About 43% of the significant SNP markers were located within known or predicted genes, including YWHAZ, KRTCAP3, TSPEAR, PIK3R4, KIF16B, PTPN3, GPRC5A, DDX47, TCF9, TPTE2, EPHA5 and NBEA genes. Our results not only confirm the results of previous reports, but also provide a suite of novel SNP markers and candidate genes associated with wool traits. Our findings will be useful for exploring the genetic control of wool traits in sheep.     

小麦矮秆突变体jm22d响应赤霉素处理的转录组学分析

为拓宽小麦矮秆遗传资源,利用γ射线辐照济麦22获得了一个赤霉素不敏感型矮秆突变体jm22d。株高相关性状调查结果及茎秆细胞学试验显示,jm22d株高为53±1.8 cm,比野生型（WT）低约20 cm。jm22d整株茎秆共有4节,比WT少一节且各节间长度显著小于WT。与WT相比,jm22d茎秆细胞长度缩短。赤霉素含量测定发现,jm22d叶片中赤霉素含量高于WT,而茎秆中赤霉素含量低于WT（P<0.01）,因此,jm22d株高降低与赤霉素转运途径出现异常有关。为了深入研究jm22d对赤霉素的响应机理,对jm22d和WT幼苗进行赤霉素处理,分别收取处理0（D0）、1（D1）和3 d（D3）的样品进行转录组学分析。结果表明,与WT相比,在jm22d中共筛选到696个上调和1 067个下调的表达基因,其中62个和349个基因在3个时间点分别表现为上调和下调表达。叶绿素含量测定表明,jm22d中叶绿素含量随赤霉素处理时间的延长而降低,聚类分析结果表明,差异表达基因主要富集在光合作用-天线蛋白（photosynthesis-antenna proteins,ko00196）、卟啉和叶绿素代谢（porphyrin and chlorophyll metabolism,ko00860）、亚油酸新陈代谢（linoleic acid metabolism,ko00591）等通路,因此赤霉素处理对jm22d体内叶绿素含量的积累具有抑制作用。通过KEGG分析在植物激素信号转导途径中挖掘到5个差异表达基因（TraesCS2B01G582300、TraesCS2B01G600800、TraesCS2B01G556600、TraesCS2B01G630000和TraesCS6B01G439600）参与生长素、细胞分裂素等激素代谢途径,这些基因在jm22d中显著下调,这可能是jm22d矮化的重要原因。研究结果为矮秆突变体矮化机制的解析提供了重要参考。     

Multi-locus genome-wide association studies reveal novel genomic regions associated with vegetative stage salt tolerance in bread wheat (Triticum aestivum L.)

Soil salinity is one of the typical abiotic stresses affecting sustainability of wheat production worldwide. In the present study, we performed a 35 K SNP genotyping assay on association panel of 135 diverse wheat genotypes evaluated for vegetative stage tolerance in hydroponics. Association analyses using five multi-locus GWAS models revealed 42 reliable QTNs for 10 salt tolerance associated traits. Among these 42 reliable QTNs, 9, 17 and 16 QTNs were associated with physiological, biomass and shoot ionic traits respectively. Novel major QTNs were identified for chlorophyll content, shoot fresh weight, seedling total biomass, Na⁺ and K⁺ concentration and Na⁺/K⁺ ratio in shoots. Further, 10 major QTNs showed significant effect on the corresponding salt tolerance traits. Gene ontology analysis of the associated genomic regions identified 58 candidate genes. The information generated in this study will be of potential value for improvement of salt tolerance of wheat cultivars using marker assisted selection.