Genetic Diversity of SSR Markers in Cultivated Hordeum vulgare L.in Qinghai Province

[Objective]The aim was to analyze genetic diversity of SSR markers in Hordeum vulgare L.in Qinghai Province and lay a foundation for screening and protecting some excellent H.vulgare cultivars.[Method]SSR markers were used to evaluate the genetic diversity of 42 cultivated H.vulgare from Qinghai Province.[Result]42 H.vulgare showed polymorphism in 7 SSR markers locus.A total of 24 alleles were identified,and the number of alleles per locus ranged from 1 to 6,with an average of 3.0.According to SSR markers polymorphism,42 H.vulgare could be divided into 4 groups,namely I,II,III and IV.[Result]The study indicated that cultivated H.vulgare from Qinghai Province is rich in genetic diversity,which will provide reference for selecting parent of H.vulgare breeding.

QTL analysis of flag leaf in barley (Hordeum vulgare L.) for morphological traits and chlorophyll content

To understand genetic patterns of the morphological and physiological traits in flag leaf of barley, a double haploid （DH） population derived from the parents Yerong and Franklin was used to determine quantitative trait loci （QTL） controlling length, width, length/width, and chlorophyll content of flag leaves. A total of 9 QTLs showing significantly additive effect were detected in 8 intervals on 5 chromosomes. The variation of individual QTL ranged from 1.9% to 20.2%. For chlorophyll content expressed as SPAD value, 4 QTLs were identified on chromosomes 2H, 3H and 6H; for leaf length and width, 2 QTLs located on chromosomes 5H and 7H, and 2 QTLs located on chromosome 5H were detected; and for length/width, I QTL was detected on chromosome 7H. The identification of these QTLs associated with the properties of flag leaf is useful for barley improvement in breeding programs.

Differential Responses of Antioxidative System to Soil Water Shortage in Barley (<i>Hordeum vulgare</i>L.) Genotypes

Drought is one of the major factors limiting the yield and quality of crops in the world. The activity of antioxidative system to tolerate the drought stress is significant in plants. In the present study, the activities and isoform profiles of catalase (CАТ), ascorbate peroxidase (APX), glutathione reductase (GR), and superoxide dismutase (SOD) were analyzed in four barley genotypes grown under soil water restriction. Drought stress caused increase in the activities of CАТ and SOD in all studied genotypes, while APX activity decreased. The total GR activity increased substantially in genotypes K 2778 and St.Garabag 7 and decreased in No. 77 local and St.Pallidum 596 genotypes under conditions of severe water stress. No detectable differences were observed in the isoenzyme pattern (the appearance of a new isoenzymes and disappearance of another one) between control plants and those subjected to soil drought. However, intensification of corresponding isoforms in electrophoretic spectra was observed in stressed barley leaves relative to watered ones. The obtained results possibly suggest that antioxidant protection in barley plants under drought conditions could be attributed mainly to SOD and CAT.

大麦HvLEC1基因的克隆及其表达特征分析

植物LEC蛋白是NF-Y转录因子的一类B亚基,在植物胚状体形成过程中起重要作用。为了研究大麦小孢子体外培养形成胚状体的机理,本研究利用RACE技术在大麦中克隆了一个新的LEC基因,该基因cDNA全长为1004 bp,开放阅读框全长为597 bp,编码198个氨基酸,其蛋白1~59位氨基酸含有LEC结构域,命名为HvLEC1。HvLEC1在大麦的根、茎、叶和小孢子培养过程中均能表达,其中小孢子培养7 d时表达量最高,且HvLEC1在大麦品系BI04中的表达量比基19高,BI04愈伤产量也比基19高,表明HvLEC1表达量和愈伤产量有相关性,受盐胁迫后HvLEC1在大麦的根中快速上调表达,提示HvLEC1可能不仅参与小孢子胚状体发生,而且参与盐胁迫响应。

几个中国大麦属物种叶绿体psbL和psbJ基因序列分析

对8份来源于中国和2份来自国外的不同大麦属物种或亚种叶绿体基因组psbL-psbJ区进行了序列测定。结果表明,10份大麦属材料的psbL-psbJ区序列长度均为365bp,且无变异位点。同时,将大麦属psbL-psbJ区序列与已报道的黑麦(Secalecereale L.)、小麦(Triticumaestivum L.)、水稻(Oryza sativa L.)和玉米(Zea mays L.)的相同序列比较,发现所有物种的psbL基因序列长度均为117bp,且无核酸变异位点;psbJ基因序列长度均为123bp,共检测到6个核酸变异位点。除水稻外,所有物种psbL和psbJ基因间区长度均为125bp,水稻psbL和psbJ基因间区为126bp。在psbL和psbJ基因间区,除水稻具有1个插入位点外,还发现8个核酸变异位点。psbL-psbJ区揭示的大麦属物种、黑麦、小麦、水稻和玉米的遗传分化距离变化范围为0.0055-0.0426。以玉米为外类群,采用邻接法进行系统发育关系分析发现,同属于小麦族的大麦属植物、黑麦和小麦间亲缘关系较近,而水稻与玉米则相对较近。

大麦cDNA-AFLP技术体系的优化及其应用

为构建适合分析大麦杂种优势的cDNA-AFLP技术体系,以4个大麦不育系、2个大麦恢复系以及按4×2配制的8个杂交种为材料,对影响大麦cDNA-AFLP技术体系的几个关键因素进行了研究。结果表明,大麦cDNA-AFLP技术按以下程序可得到多态性好的清晰条带:使用百泰克公司的TRIpure Reagent总RNA提取试剂可提取到高质量的叶片总RNA;用TaKaRa的M-MLV RTase反转录合成双链cDNA;用MseⅠ和EcoRⅠ酶切6h,再用T4连接酶15℃连接接头18h;预扩PCR反应的dNTP终浓度为0.2mmol.L-1,扩增20循环效果最好;预扩产物稀释20倍后进行选择性扩增,最终用6%的变性聚丙烯酰胺凝胶进行电泳分离条带。利用该cDNA-AFLP体系对参试杂交种及其亲本的谱带类型进行了分析,亲本与杂种之间共扩增出7种类型的谱带,分别是共同表达型(P1P2F1)、P1表达型(P1)、P2表达型(P2)、F1表达型(F1)、P2F1表达型(P2F1)、P1F1表达型(P1F1)和P1P2表达型(P1P2)。其中差异谱带类型可分为四种表达模式:(1)单亲显性表达型,包括P2F1表达(P2F1)和P1F1表达(P1F1);(2)单亲沉默表达型,包括仅P1表达(P1)和仅P2表达(P2);(3)杂种上调表达型,即仅F1表达(F1);(4)杂种下调表达型,即仅P1P2表达(P1P2)。

大麦种子萌发lncRNA-miRNA-mRNA差异表达分析及ceRNA调控网络构建

为解析长链非编码RNA(lncRNA)、microRNA(miRNA)和mRNA如何在ceRNA互作模式下调控大麦种子萌发,揭示大麦(Hordeum vulgare L.)种子萌发转录水平分子调控机制,基于大麦种子萌发相关的RNA-Seq数据,共鉴定出6447个差异表达的mRNA,661个差异表达的lncRNA,310个差异表达的miRNA;对差异表达的mRNA和lncRNA进行加权基因共表达网络(WGCNA)分析,各鉴定到1个与大麦种子萌发高度相关的共表达模块。模块内相关基因涉及水分响应、胚发育等生物学过程,以及MAPK信号转导、植物激素信号转导等信号通路。通过对lncRNA/mRNA和miRNA进行靶基因预测,得到168个miRNA-mRNA靶基因对以及310个miRNA-lncRNA靶基因对,进而构建了包含2个子网络的ceRNA调控网络,发现有16个miRNA、38个lncRNA以及18个mRNA可能具有ceRNA功能。最后,利用qRT-PCR验证了2个关系对:HORVU0Hr1G039470-miR2611-MSTRG.21252以及HORVU6Hr1G031480-miR1858-MSTRG.23227。

NaCl对大麦硝态氮吸收动力学特征的影响

采用营养液培养方法研究了不同盐分和氮水平对大麦‘鉴四’(Hordeum vulgare L.)生长及硝态氮(NO3?-N)吸收动力学参数特征的影响。结果表明:不同浓度NO3?-N与Na Cl预处理后,大麦对NO3?吸收符合离子吸收动力学模型,其吸收动力学参数表现为NO3?预处理浓度增加后,Vmax增大,Km值增加,但增加的幅度不一致。对高亲和力系统来说,所有预处理大麦的NO3?-N吸收曲线均符合Michaelis-Menten方程的描述。1 mmol?L?1NO3?-N预处理中,120 mmol?L?1 Na Cl预处理比1 mmol?L?1 Na Cl预处理显著提高了NO3?-N的吸收速率;10 mmol?L?1NO3?-N预处理时,120 mmol?L?1 Na Cl预处理与1 mmol?L?1 Na Cl预处理无显著性差异。表明低氮环境下生长的大麦,其根系NO3?-N高亲和力系统受Na Cl影响较大。对低亲和力系统来说,所有预处理大麦的NO3?-N吸收曲线均符合Michaelis-Menten方程的描述。在1 mmol?L?1 NO3?-N预处理中,与1 mmol?L?1 Na Cl预处理相比,120 mmol?L?1 Na Cl胁迫的大麦NO3?-N吸收速率显著提高;10 mmol?L?1 NO3?-N预处理中,120 mmol?L?1 Na Cl与1 mmol?L?1 Na Cl处理相比,降低了大麦的NO3?-N吸收速率。表明低氮环境下生长的大麦,盐胁迫对其根系低亲和力系统有促进作用,而高氮环境下生长的大麦,盐胁迫对其低亲和力系统的NO3?-N吸收没提高作用。

HPLC法同时测定大麦苗中5种成分

目的建立HPLC法同时测定大麦苗Hordeum vulgare L.中大麦黄苷、皂草苷、异荭草素-7-O-[6-阿魏酰基]-葡萄糖苷、异牡荆素-7-O-[6-阿魏酰基]-葡萄糖苷、异荭草素-2″-O-[6-阿魏酰基]-葡萄糖苷的含量。方法大麦苗30%乙醇提取物的分析采用Waters Atlantis■ T3色谱柱(4.6 mm×250 mm,5μm);流动相0.2%甲酸-乙腈,梯度洗脱;体积流量1.0~1.5 mL/min;柱温30℃;检测波长345 nm。结果5种成分在各自范围内线性关系良好(r>0.9996),平均加样回收率98.12%~102.40%,RSD 2.12%~2.65%。结论该方法准确稳定,重复性好,可用于大麦苗的质量控制。

Evaluation of Chlorophyll Content and Fluorescence Parameters as Indicators of Drought Tolerance in Barley

Drought is a major abiotic stress that severely affects food production worldwide. Agronomic and physiological traits associated with drought tolerance are suitable indicators for selection of drought tolerance genotypes to reduce the impact of water deficit on crop yield in breeding program. The objective of this study was to identify indicators related to drought tolerance through analysis of photosynthetic traits in barley （Hordeum vulgare L.）. These traits included chlorophyll content, initial fluorescence （Fo）, maximum primary yield of photochemistry of photosystem Ⅱ （Fv /Fo） and maximum quantum yield of photosystem Ⅱ （Fv/Fm）. Four genotypes （Tadmor, Arta, Morocco9-75 and WI2291） variable in drought tolerance were used to investigate the correlation between these traits and drought tolerance. The results reflected that all of these traits were affected negatively in the four genotypes at different levels of post-anthesis drought stress, but the decrease in drought tolerant genotypes was much less than that of drought sensitive genotypes. The results further revealed that the components of the photosynthetic apparatus could be damaged significantly in drought sensitive genotypes, while drought tolerant genotypes were relatively less affected. On the other hand, the values of chlorophyll content, Fo, Fv/Fo and Fv/Fm in drought tolerance genotypes were significantly higher than those in drought sensitive genotypes under drought stress. It was concluded that chlorophyll content, Fo, Fv/Fo and Fv/Fm could be considered as reliable indicators in screening barley germplasm for drought tolerance.

大麦TILLING群体的构建及EDR1基因突变的检测(英文)

该研究选用栽培大麦品种‘西引2号’,通过化学诱变剂甲基磺酸乙酯(ethyl methane sulfonate,EMS)处理,创建了3 750个M1及2 012个M2突变株系。结果表明:在M2中,有115株表现黄化,98株表现矮化,101株表现极早或极晚抽穗,其他35株表现分蘖少、叶片小或者不育等性状,所有突变株系占M2群体的17.35%;M2的基因组DNA用于突变频率的研究,用优化的CELⅠ酶切体系对M2株系进行了EDR1基因突变体筛选,获得了EDR1基因的3个突变体,其中一个发生在外显子区域,导致了氨基酸Pro到Ser的转换,突变频率平均为每661kb一个点突变,这与前人研究的突变频率基本在一个范围内。该研究创建了大麦品种‘西引2号’的TILLING筛选群体,并用于EDR1突变体的筛选。为大麦性状研究提供了不同的思路,搭建了新的资源和技术平台。

Influence of Aluminum and Cadmium Stresses on Mineral Nutrition and Root Exudates in Two Barley Cultivars

A hydroponic experiment was carried out to study the effect of aluminum （Al） and cadmium （Cd） on Al and mineral nutrient contents in plants and Al-induced organic acid exudation in two barley varieties with different Al tolerance. Al- sensitive cv. Shang 70-119 had significantly higher Al content and accumulation in plants than Al-tolerant cv. Gebeina, especially in roots, when subjected to low pH （4.0） and Al treatments （100 μmol L^-1 Al and 100 μmol L^-1 Al ＋1.0 μmol L^-1 Cd）. Cd addition increased Al content in plants exposed to Al stress. Both low pH and Al treatments caused marked reduction in Ca and Mg contents in all plant parts, P and K contents in the shoots and leaves, Fe, Zn and Mo contents in the leaves, Zn and B contents in the shoots, and Mn contents both in the roots and leaves. Moreover, changes in nutrient concentrations were greater in the plants exposed to both Al and Cd than in those exposed only to Al treatment. A dramatic enhancement of malate, citrate, and succinate was found in the plants exposed to 100 μmol L^-1 Al relative to the control, and the Al-tolerant cultivar had a considerable higher exudation of these organic acids than the Al-sensitive one, indicating that Al-induced enhancement of these organic acids is very likely to be associated with Al tolerance.

Alleviation of Al Toxicity in Barley by Addition of Calcium

The potential mechanism by which Ca alleviates Al toxicity was investigated in barley seedlings. It was found that 100 Al-alone treatment inhibited barley plant growth and thereby reduced shoot height and root length, and dry weights of root, shoot and leaf; promoted Al accumulation but inhibited Ca absorption in plant tissues; and induced an increase in the activities of superoxide dismutase （SOD）, catalase （CAT）, and peroxidase （POD） and in the level of lipid peroxidation （MDA content） in leaves. Except for the increase in Ca concentration in plant tissues, treatment with 0.5 mM Ca in the absence of Al had less effect on the above-mentioned parameters, compared with the control. Addition of Ca efficiently reduced Al toxicity, which is reflected by the promotion of plant growth, reduction in Al concentration and MDA content, increase in Ca concentration and in SOD, POD, and CAT activities compared with the Al-alone-treatment; with increase in Ca level （3.0 raM）, the ameliorative effect became more dominant. This indicated that the alleviation of aluminum toxicity in barley seedlings with Ca supplementation could be associated with less absorption of Al and the enhancement of the protective ability of the cell because of increased activity of the antioxidative enzyme.

Effects of Waterlogging on Photosynthesis and Antioxidant Enzyme Activities of Six Barley Genotypes with Different Waterlogging Tolerance

A field experiment was carried out to study genotypic difference in the effect of waterlogging on photosynthesis, chlorophyll content and antioxidative enzyme activities in barley. Waterlogging caused a rapid decline in net photosynthetic rate (Pn) and stomatal conductance (gs), and little change in chlorophyll content during early days of the treatment. A dramatic increase in malondialdehyde (MDA) content, superoxide dismutase (SOD) and peroxidase (POD) in waterlogged plants in the early days of the experiment was found, indicating the occurrence of oxidative stress in barley plants exposed to waterlogging. There was a highly significant difference in the changed extent of all these parameters among genotypes. Franklin and Yongjiahong Liuleng Damai, which were relatively sensitive to waterlogging in terms of growth, photosynthesis and chlorophyll content, accumulated much more MDA than the other two relatively tolerant genotypes (93-3143 and QS). After removal of waterlogging, the genotypic difference became much greater in recovering of these examined parameters. Yongjiahong Liuleng Damai showed higher recovery, while Franklin only recovered to 50% of the control at the 14 day after waterlogging removal. It may be concluded that it is the difference in anti-oxidative stress caused by waterlogging that account for the major difference in photosynthesis among barley genotypes.

Genotypic and Environmental Variations of Arabinoxylan Content and Endoxylanase Activity in Barley Grains

Arabinoxylan （AX） content in barley grains is an important quality determinant when barley is used as raw material of malt or beer production. The cultivar and environmental variations of total arabinoxylan （TAX）, water extractable arabinoxylan （WEAX） and endoxylanase activity （EA） were investigated using eight barley cultivars growing at seven locations with diverse environmental conditions. The results showed that both barley cultivar and location significantly affected the TAX, WEAX and EA levels, but the variations of TAX content and EA were mainly attributed to cultivar, while the impact of location on WEAX content was greater than that of cultivar. Correlation analysis indicated that TAX was significantly correlated to WUAX.

Genotypic Difference in Plant Growth and Mineral Composition in Barley Under Aluminum Stress

Four barley genotypes (Tiantaiyangdamai, Xiyin2, Mimaill4 and Tai94-Ce6) were exposed to 0, 50, 100, and 150μM of Al-containing solution with pH 4.5, to determine the differences in growth inhibition , Al concentration and accumulation and mineral composition among genotypes. The results showed that Mimaill4 and Tai94-Ce6 had significantly higher Al concentration and accumulation than Tiantaiyangdami and Xiyin2, especially in roots, and the growth traits including root and shoot dry weights, shoot height, root length and tillers per plant were more inhibited in the former two genotypes. Al treatments caused a significant reduction of N, P, K, Ca, Mg and Mn content in both roots and shoots, of Cu in shoots; and a significant increase in Fe and Zn content in both roots and shoots, of Cu in roots. The changed rates of mineral content caused by Al treatments, in terms of the content in 150μM Al divided by the content in the control, differed significantly among four genotypes. Two Al-sensitive genotypes, Mimaill4 and Tai94-Ce6 had much greater changes in mineral content than other two Al-tolerant genotypes Tiantaiyangdamai and Xiyin2 when subjected to Al stress in comparison with the control. It is indicated that the Al-tolerant genotype is characterized by less uptake and accumulation of Al in roots and smaller disorders in mineral metabolism and ion homeostasis.

Variation of Grain and Malt Qualities in Barley as Affected by Cultivars and Environments

Inferior and unstable quality of malt barley in China has limited its wide use in malting and brewing industries. In this research, eight two-rowed barley cultivars were planted at 7 locations with different ecological conditions in southern China, to investigate the cultivar and environmental effect on grain and malt qualities. The results showed that grain protein content differed dramatically among locations, but there were no significant differences among cultivars. For four malt qualities including diastatic power, wort viscosity, Kolbach index and malt extract, significant differences were found among cultivars and locations, except for diastatic power among cultivars. Coefficients of variance(CV)caused by location were greater than those caused by cultivar for each quality parameter, especially for diastatic power and Kolbach index, indicating the predominant influence of environment on malt quality. The analysis showed that grain protein content was positively related to diastatic power and wort viscosity, and negatively to malt extract. Kolbach index was positively related to malt extract, and negatively to diastatic power and wort viscosity. Diastatic power showed positive and negative correlation with wort viscosity and malt extract, respectively.

Protein and hordein fraction content in barley seeds as affected by sowing date and their relations to malting quality

The effect of sowing date on grain protein, hordein fraction content and malting quality of two-rowed spring barley was investigated by using ten commercial cultivars with different grain protein content and the relationships among these traits were examined. The results showed that grain protein content and B hordein content increased as the sowing date postponed and were significantly affected by sowing date, while C and D hordein contents were less influenced by sowing date. There were significant differences in grain protein and hordein fraction content among the ten cultivars. The coefficient of variation of D hordein content was much larger than that of B and C hordein contents, suggesting its greater variation caused by different sowing dates. Beta-amylase activity and diastatic power were also significantly affected by sowing date, with malt extract being less affected. Significant differences in measured malt quality were found among the ten cultivars. Grain protein was significantly correlated with B hordein and malt extract positively and negatively, respectively. There was no significant correlation between beta-amylase activity or diastatic power and grain protein content. B hordein was negatively and significantly correlated with malt extract, but no significant correlations between C hordein, D hordein and malting quality traits.

Cultivar and Environmental Effects on p-glucanase Activity in Both Barley Grain and Malt and Its Function in β-glucan Degradation

Eight two-rowed barley cultivars were grown at seven locations in the southern winter-barley zone of China. Mean grainβ-glucanase activity ranged from 39.89 U kg-1 for Suyin2 to 49.75 U kg-1 for Xi-umai3 in 8 cultivars grown at 7 locations, and from 38. 74 U kg-1 in Zhengzhou to 57. 96 U kg-1 in Putian among 7 locations on an average of all cultivars. Correspondingly, mean malt β-glucanase activity of 8 cultivars ranged from 313.33 U kg-1 for ZAU3 to 489. 89 U kg-1 for Daner Barley, and of 7 locations from 330.40 U kg-1 in Yancheng to 418. 24 U kg-1 in Putian. There were significant differences among cultivars and locations in maltβ-glucanase activities. The locations showed much larger variation in maltβ-glucanase activities than cultivars. The reduction of total β-glucan content after malting varied in both cultivars and locations, with a mean of 78.31%. The analysis of correlations showed that maltβ-glucan content was significantly positively and negatively correlated with grain β-glucan content and malt β-glucanase activity, respectively, and malt β-glucanase activity was significantly positively correlated with grain β-glucanase activity.

NaCl胁迫对青稞种子萌发及幼苗生长的影响

以西藏自治区5个青稞(Hordeum vulgare L. var. nudum Hook. f.)品种为试验材料,研究不同NaCl浓度对青稞种子萌发及幼苗生长的影响,并且对不同青稞品种耐盐性进行评价分析。结果表明,NaCl浓度为0.1%时对青稞萌发及幼苗生长具有一定的促进作用;当浓度增至0.5%时,种子的发芽势、发芽率、活力指数以及幼苗的根长、芽长、根鲜重均显著下降;当浓度为1.0%时,隆子黑青稞与苟芝紫青稞已丧失发芽能力。总体来看,5个青稞品种中,喜拉22耐盐性最好,其次是藏青320、藏青2000、苟芝紫青稞,隆子黑青稞耐盐性最弱。