Sequence Polymorphisms and Phylogenetic Relationships of hina Gene in Wild Barley from Tibet, China

The hina gene encodes a HINA protein in seeds of barley （Hordeum vulgare）, which was known to affect the grain hardness. 171 hina gene sequences from Tibetan wild barley accessions and worldwide were characterized. Across 1 452 nucleotides of 171 hina genes, 152 SNPs were detected, giving an average frequency of one SNP per 9.5 bases. There were 93 singleton variable sites （the nucleotide polymorphism only observed in a single accession）, 59 polymorphic sites （the polymorphisms found in two or more accessions） and 8 indels. A total of 18 haplotypes were defined, and most of the barley accessions shared one gene haplotype. H. spontaneum had a wider haplotype distribution. Through the analysis of median-joining network of the 18 haplotypes, 4 haplotype groups were found, which were testified by neighbor-joining tree based on the complete sequence alignment. Extremely low level of hina gene diversity was observed in Tibetan wild barley accessions, indicating that Tibet is unlikely a center of origin for cultivated barley.     

Accumulation of Protein Fractions during Grain Filloing of Wheat Genotypes Differing in Protein Content and Baking Quality

The accumulation of protein fractions was analyzed on developing and mature wheat grains of three cultivars differing in protein content and baking quality.There was a slight difference in the accumulation of cytoplasmic proteins in the cultivars used.The high yield but low protein cultivar showed a consistent decline of protein content during grain filling but the high-protein cultivars increascd their psotein contant after 25 days post-anthesis.The accumulation of storage proteins was different from that of cytoplasmic protein.and there were also cultivar variations,However,all cultivars reached their.Maximum-synthesizing capacity for storage proteins at maturity.The relationship between the protein fractions or their ratio and baking quality was also discussed.     

Accumulation of Protein Fractions during Grain Filling of Wheat Genotypes Differing in Protein Content and Baking Quality

The accumulation of protein fractions was analyzed on developing and mature wheat grains of three cultivars differing in protein content and baking quality. There was a slight difference in the accumulation of cytoplasmic proteins in the cultivars used. The high yield but low protein cultivar showed a consistent decline of protein content during grain filling but the high - protein cultivars increased their psotein contant after 25 days post-anthcsis. The accumulation of storage proteins was different from that of cytoplasmic protein, and there were also cultivar variations. However, all cultivars reached their, maximum-synthesizing capacity for storage proteins at maturity. The relationship between the protein fractions or their ratio and baking quality was also discussed.     

Influence of different nitrogen application on flour properties,gluten properties by HPLC and end-use quality of Korean wheat

This study was performed to identify how the different levels of nitrogen application affected the variances of gluten properties and end-use qualities and the differences of variances among Korean wheat cultivars. Protein and dry gluten content, SDS sedimentation volume and water absorption of Mixolab increased as nitrogen application increased. This ratio of the increase was higher in Korean wheat cultivars for bread than in Korean wheat cultivars for noodles and cookies. The proportion of(α+β)-gliadin measured by reversed-phase high-performance liquid chromatography(RP-HPLC) increased, but the proportion of ω-and γ-gliadin decreased as the protein content increased. The Korean wheat cultivars for bread showed a high proportion of(α+β)-gliadin increase, the Korean cultivars for noodles had a high proportion of γ-gliadin decrease and the Korean wheat cultivars for cookies had a high proportion of ω-gliadin decrease. However, there was no variation of the component in the proportion of glutenin component measured by RP-HPLC, even though the protein content was increased, but all of the protein fractions measured by size exclusion(SE)-HPLC were increased. The soluble monomeric protein showed a high proportion of Korean wheat cultivars for bread by the increase of protein content. Bread loaf volume increased by the increase of protein content but there were no variances in the ratio of increase among Korean wheat cultivars. The cookie diameter decreased with the increase of protein content, and this ratio of decrease was the highest in Korean wheat cultivars for cookies. The hardness of cooked noodles also increased by the increase of protein content but there were no variations in springiness and cohesiveness. The decrease proportion of ω-gliadin affected the increase of bread loaf volume, the hardness of cooked noodles, and the decrease of cookie diameter.     

Influence of protein characteristics and the proportion of gluten on end-use quality in Korean wheat cultivars

The effects of protein characteristics and the proportion of gluten on end-use quality in 13 Korean wheat cultivars for three years were verified in this study. Year, cultivar, and the interaction between the year and the cultivar influenced protein characteristics, the proportion of gluten except for γ-and ω-gliadin using RP-HPLC(reversed-phase high-performance liquid chromatography), and end-use quality. Protein characteristics and the proportion of gluten in Korean wheat cultivars were between those of Australian standard white(ASW) and hard wheat(AH). Korean wheat cultivars exhibited a higher average α+β gliadin proportion than imported wheat, a γ-gliadin proportion similar to that of dark northern spring wheat, and the same ω-gliadin proportion as AH. They showed a bread-loaf volume intermediate between those of ASW and AH and a texture of cooked noodles similar to that of soft white wheat, but less springiness than imported wheat. The cookie diameter of Korean wheat cultivars was similar to that of hard red winter wheat. There was a correlation between bread-loaf volume and protein characteristics, except for the protein content in Korean wheat cultivars. Springiness and cohesiveness of cooked noodles were not correlated with protein characteristics, while hardness was correlated with the protein content and water absorption of a mixograph. Cookie diameter was negatively correlated with the sodium dodecyl sulfate(SDS) sedimentation volume and water absorption of a mixograph. The end-use quality was not correlated with any proportion of gluten composition. Principal component analysis(PCA) showed that the proportion of gluten was not related to the quality of the bread(both PCs, 81.3%), noodle(77.7%), and cookie(82.4%). PCA explained that Keumkang is suitable for superior bread, while Uri is good for cooked noodles and cookies.     

Postulation of Seedling Resistance Genes in 20 Wheat Cultivars to Yellow Rust （ Puccinia striiformis f. sp. triticl）

The gene postulation, cluster analysis and pedigree analysis of 20 wheat cultivars to 28 physiological races of yellow rust （Puccinia striiformis） were conducted under greenhouse conditions, The spectrum of their resistance were compared each other. None of the 20 cultivars were resistant to all the test pathogens. The cultivars containing resistance genes Yr5 and Yr24 were not find and genes Yr8, Yrl9 and Yr27 could not be postulated from cultivars tested due to the susceptibility to all isolates used. A total of 6 probale seedling yellow rust resistance genes or gene combinations （Yrl, Yr2, Yr2 ＋ YrHVII, Yr3 ＋ unknown, Yr3 ＋ Yr4, YrAlba） were postulated in the wheat cultivars （Atou, Flanders, Maris Huntsman, Bouquet, Holdfast, Elite Lepeuple, and Vilmorin 27）. The gene combination Yr2＋YrHVII with the highest frequency （35%） was present in 7 cultivars. The Yr genes present in some wheat cultivars could not be postulated because of non-matching virulence combinations with any of known genes. Cluster result showed that Yr2 and Yr3 are the most important genes in the cultivars. The 13 cultivars are believed to have the pedigree of Noe, which was selected from South Russian wheat. These results will be useful for wheat breeding and provide information about genetic control of wheat yellow rust.     

The Effects of Dwarfing Genes （Rht-B1b,Rht-D1b,and Rht8） with Different Sensitivity to GA3 on the Coleoptile Length and Plant Height of Wheat

Understanding the effects of wheat dwarfing genes on the coleoptile length and plant height is crucial for the proper utilization of dwarfing genes in the improvement of wheat yield. Molecular marker analysis combined with pedigree information were used to classify wheat cultivars widely planted in major wheat growing regions in China into different categories based on the dwarfing genes they carried. The effects of the dwarfing genes with different sensitivity to gibberellins （GA3） on the coleoptile length and plant height were analyzed. Screening of 129 cultivars by molecular marker analysis revealed that 58 genotypes of wheat contained the dwarfing gene Rht-B1b, 24 genotypes of wheat contained Rht-D1b gene and 73 genotypes of wheat possessed Rht8 gene. In addition, among these 129 cultivars, 35 genotypes of wheat cultivars contained both Rht-B1b and Rht8 genes and 16 genotypes of wheat cultivars contained both Rht-D1b and Rht8 genes. Wheat cultivars with the dwarfing genes Rht-B1b or Rht-D1b were insensitive to GA3, while the cultivars with the dwarfing gene Rht8 were sensitive to GA3. Most of the wheat genotypes containing combination of Rht8 gene with either Rht-B1b or Rht-D1b gene were insensitive to GA3. The plant height was reduced by 24.6, 30.4, 28.2, and 32.2%, respectively, for the wheat cultivars containing Rht-B1b, Rht-D1b, Rht-B1b ＋ Rht8, and Rht-D1b ＋ Rht8 genes. The plant height was reduced by 14.3% for the wheat cultivar containing GA3-sensitive gene Rht8. The coleoptile length was shortened by 25.4, 31.3, 28.4 and 31.3%, respectively, in the wheat cultivars containing Rht-B1b, Rht-D1b, Rht-B1b ＋Rht8 and Rht-D1b ＋ Rht8 genes, while the coleoptile length was shortened only by 6.2% for the wheat cultivar containing Rht8 gene. We conclude that GA3-insensitive dwarfing genes （Rht-B1b and Rht-D1b） are not suitable for the wheat improvement in dryland because these two genes have effect on reducing both plant height and coleoptile length. In contrast, GA3- sensitive dwarfing gene （Rht8） is a relatively ideal candidate for the wheat improvement since it significantly reduces the plant height of wheat, but has less effect on the coleoptile length.     

Study on Accumulation Rule of Carbohydrates in Soybean （Glycine max（L.） Merrill）

Three soybean cultivars, Hefeng25 （HF25）, Suinongl4 （SN14） and Longxuanl （LX1） were used as materials and the accumulation features of carbohydrates were studied in leaves and stems. The results showed that the soluble sugar content in leaves and stems appeared the same trend of higher-lower-higher in three varieties. The changes of sucrose content in leaves appeared the same rule with that of soluble sugar, that is higher-lower-higher, while presented a rising trend in stems. The content of starch in leaves varied as a change of double apexes. The content of starch in stems appeared a change of single apex. In the same stage, there were some differences among carbohydrates. The content of soluble sugar and sucrose in SN14 was higher, and the starch content was lower.     

Study on the Change of GAs,ABA Level and ABA／GAs Ratio in the Embryos of Spring Wheat and Winter Wheat During Vernalization

Spring wheat(cv.7742)and winter wheat (cv.Nongda 015) were vernalized by seed chilling,and the changes of GAs,ABA content and GAs/ABA ration in embryo wer investigated.The differences between cultivars,treatments and variety-treatment inleractions were significant for both GAs and ABA.The level of GAs and ABA as well as GAs/ABA ratio were hardly changed by seed chilling in spriug wheat Howerve,in winter wheat,the GAs content decreased much less compared with ABA that reduced sharply after chilling.It showed that the GAs/ABA ratio in winter wheat was increased remarkably(4.55 times)than in spring wheat.It was suggested that the absolute GAs Ievel was independent of the vernalization effect because there was a higher GAs Level before and after seed chilling in winter wheat when compared with spring wheat,One of the vemalization effects was the decline of ABA level that led to the increase of GAs/ABA ratio.     

Expression analysis,single nucleotide polymorphisms within SIRT4 and SIRT7 genes and their association with body size and meat quality traits in Qinchuan cattle

Silent information regulator 2(Sir2) proteins, or sirtuins, are nicotine adenine dinucleotide(NAD)-dependent deacetylases that connect metabolism with longevity in lower organisms. In mammals, there are seven Sir2 homologs, namely, silent information regulators(SIRT1–7). SIRT4 and SIRT7 genes play a crucial role in regulating lipid metabolism, cellular growth and metabolism. This suggests that they are potential candidate genes for affecting body size and meat quality traits in animals. Hence, this study aimed to detect genetic variations of both SIRT4 and SIRT7 bovine genes in Qinchuan cattle, and to evaluate the effect of these variations on economically important body size and meat quality traits. Expression analysis using quantitative real-time PCR(q PCR) indicated that SIRT4 and SIRT7 were broadly expressed in all thirteen studied tissues. The expression of SIRT4 was higher in liver, muscle, and in subcutaneous fat tissue. In the case of SIRT7, the expression was higher in lung, abomasum, and subcutaneous fat. Using DNA sequencing, a total of three single nucleotide polymorphisms(SNPs) were identified within SIRT4 and SIRT7 genes in 468 Qinchuan cattle. These included one novel SNP within 3′ untranslated regions(UTR) of SIRT4(SNP1: g.13915AG) and two novel synonymous substitutions in SIRT7(SNP2: g.3587CT and SNP3: g.3793TC). Statistical analyses indicated that all three SNPs could significantly influence some body size and meat quality traits in Qinchuan cattle. These novel findings will provide a background for application of bovine SIRT4 and SIRT7 genes in the selection program of Chinese cattle.     

Identification of the resistance gene to powdery mildew in Chinese wheat landrace Baiyouyantiao

Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is one of the most damaging diseases to wheat in the world. The cultivation of resistant varieties of wheat is essential for controlling the powdery mildew epidemic. Wheat landraces are important resources of resistance to many diseases. Mapping powdery mildew resistance genes from wheat landraces will promote the development of new varieties with disease resistance. The Chinese wheat landrace Baiyouyantiao possesses characteristic of disease resistance to powdery mildew. To identify the resistance gene in this landrace, Baiyouyantiao was crossed with the susceptible cultivar Jingshuang 16 and seedlings of parents and F₁, BC₁, F₂, and F_2:3 were tested with Bgt isolate E09. The genetic results showed that the resistance of Baiyouyantiao to E09 was controlled by a single recessive gene, tentatively designated PmBYYT. An Illumina wheat 90K single-nucleotide polymorphism (SNP) array was applied to screen polymorphisms between F₂-resistant and F₂-susceptible DNA bulks for identifying the chromosomal location of PmBYYT. A high percentage of polymorphic SNPs between the resistant and susceptible DNA bulks was found on chromosome 7B, indicating that PmBYYT may be located on this chromosome. A genetic linkage map of PmBYYT consisting of two simple sequence repeat markers and eight SNP markers was developed. The two flanking markers were SNP markers W7BL-8 and W7BL-15, with genetic distances of 3 and 2.9 cM, respectively. The results of this study demonstrated the rapid characterization of a wheat disease resistance gene and SNP marker development using the 90K SNP assay. The flanking markers of gene PmBYYT will benefit marker-assisted selection (MAS) and map-based cloning in breeding wheat cultivars with powdery mildew resistance.     

大麦糯性相关基因Wx单核苷酸多态性分析

【目的】大麦Wx是控制直连淀粉合成相关的糯性基因,研究大麦糯性相关基因Wx单核苷酸多态性,并分析其与籽粒直链淀粉含量的关系。【方法】以2个国外糯大麦品种为对照,对30份高、中、低直链淀粉含量的中国大麦进行Wx的克隆和测序,分析Wx的单核苷酸多态性(SNP)及其与籽粒直链淀粉含量之间的关系。【结果】在对32个大麦品种的核苷酸序列多态性鉴定中,共检测到了169个多态性位点,平均每26bp检测到一个多态性位点。在所有检测到的多态性位点中,包括143个SNP和26个InDel,二者的频率分别为1/310和1/169。Wx的内含子1、3、5、8区,外显子2、5和5′-UTR及3′-UTR区域为变异富集区,其它区域变异较小。外显子2和内含子1区域所承受的选择压力较小。单倍型分析表明,第1种单倍型中包括所有低直链淀粉含量的材料。【结论】大麦Wx的多态性与直链淀粉含量之间存在明显的对应关系。     

Comparative Analysis of Hina Gene Sequences in Wild （Hordeum spontaneum） and Cultivated （H. vulgare） Barleys

The Hina gene is one of the two known Hin genes for hardness, and its RNA expression is correlated with grain hardness and dry matter digestibility variation. In this study, only one clone of Hina gene was obtained from one barley accession. A total of 121 Hina gene sequences were isolated from 121 wild barley {Hordeum spontaneum) accessions in Israel, Iran, and Turkey, and then their molecular characteristics were compared with 97 Hina gene sequences from 74 cultivated barley (H. vulgäre) lines in Europe and 23 landrace (H. vulgäre) with global distribution and other 26 Hina gene sequences from cultivated barleys (H. vulgäre) with unknown global distribution. C/s-acting regulatory element (CARE) searching revealed that there were different types of regulatory element for the Hina gene in wild and landrace/cultivated barleys. There were six consistent cw-acting binding sites in wild and landrace/cultivated barleys, whereas 8 to 16 inconsistent TATA-boxes were observed. In addition, three special elements (E2Fb, Spl, and boxS) were only observed in wild barley, while one (AT 1 -motif) was only found in landrace/cultivated barley. Forty-four deduced amino acid sequences of HINA from wild and landrace/cultivated barleys were obtained by deleting repetitive amino acid sequences, and they were clustered into two groups on the basis of Neighbor-Joining analysis. However, there was no obvious difference in the amino acid sequences of HINA between wild and landrace/cultivated barleys. Comparing to protein secondary structure of wheat PINA, it was indicated that HINA also existed a signal peptide. In addition, HINA was a hydrophilic protein on the basis of the protein properties and composition.     

Genetic diversity analysis and GWAS reveal the adaptive loci of milling and appearance quality of japonica rice (Oryza sativa L.) in Northeast China

Milling and appearance quality are important contributors to rice grain quality. Abundant genetic diversity and a suitable environment are crucial for rice improvement. In this study, we investigated the milling and appearance quality-related traits in a panel of 200 japonica rice cultivars selected from Liaoning, Jilin and Heilongjiang provinces in Northeast China. Pedigree assessment and genetic diversity analysis indicated that cultivars from Jilin harbored the highest genetic diversity among the three geographic regions. An evaluation of grain quality indicated that cultivars from Liaoning showed superior milling quality, whereas cultivars from Heilongjiang tended to exhibit superior appearance quality. Single- and multi-locus genome-wide association studies (GWAS) were conducted to identify loci associated with milling and appearance quality-related traits. Ninety-nine significant single-nucleotide polymorphisms (SNPs) were detected. Three common SNPs were detected using the mixed linear model (MLM), mrMLM, and FASTmrMLM methods. Linkage disequilibrium decay was estimated and indicated three candidate regions (qBRR-1, qBRR-9 and qDEC-3) for further candidate gene analysis. More than 300 genes were located in these candidate regions. Gene Ontology (GO) analysis was performed to discover the potential candidate genes. Genetic diversity analysis of the candidate regions revealed that qBRR-9 may have been subject to strong selection during breeding. These results provide information that will be valuable for the improvement of grain quality in rice breeding.     

QTL-seq analysis of the seed size trait in grape provides new molecular insights on seedlessness

Seedlessness in grape(Vitis vinifera) is an important commercial trait for both the fresh and drying markets. However, despite numerous studies, the mechanisms and key genes regulating grape seedlessness are mostly unknown. In this study, we sequenced the genomes of the V. vinifera seeded cultivar ‘Red Globe’, the seedless cultivar ‘Centennial Seedless’, and the derived hybrids. Nonsynonymous single nucleotide polymorphisms(SNPs) were identified by genome sequencing and analyzed using published ...     

Sequence Polymorphisms and Phylogenetic Relationships of hina Gene in Wild Barley from Tibet, China

The hina gene encodes a HINA protein in seeds of barley (Hordeum vulgare), which was known to affect the grain hardness. 171 hina gene sequences from Tibetan wild barley accessions and worldwide were characterized. Across 1 452 nucleotides of 171 hina genes, 152 SNPs were detected, giving an average frequency of one SNP per 9.5 bases. There were 93 singleton variable sites (the nucleotide polymorphism only observed in a single accession), 59 polymorphic sites (the polymorphisms found in two or more accessions) and 8 indels. A total of 18 haplotypes were defined, and most of the barley accessions shared one gene haplotype. H. spontaneum had a wider haplotype distribution. Through the analysis of median-joining network of the 18 haplotypes, 4 haplotype groups were found, which were testified by neighbor-joining tree based on the complete sequence alignment. Extremely low level of hina gene diversity was observed in Tibetan wild barley accessions, indicating that Tibet is unlikely a center of origin for cultivated barley.     

Genome-wide association study for starch content and constitution in sorghum (Sorghum bicolor (L.) Moench)

Starch is the most important component in endosperm of sorghum grain. Usually, two types of starch are present: amylose (AM) and amylopectin (AP). The levels of AM and AP contents play a significant role in the appearance, structure, and quality of sorghum grains and in marketing applications. In the present study, a panel of 634 sorghum (Sorghum bicolor (L.) Moench) accessions were evaluated for starch, AM, and AP contents of grain, which included a mini core collection of 242 accessions from the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) in India, and 252 landraces and 140 cultivars from China. The average starch content was 67.64% and the average AM and AP contents were 20.19 and 79.81%, respectively. We developed a total of 260 000 high-confidence single nucleotide polymorphism (SNP) markers in the panel of 634 accessions of S. bicolor using specific locus amplified fragment sequencing (SLAF-seq). We performed genome-wide association studies (GWAS) of starch, AM, and AM/AP of grain and SNP markers based on a mixed linear model (MLM). In total, 70 significant association signals were detected for starch, AM, and AM/AP ratio of grain with P<4.452×10^–7, of which 10 SNPs were identified with significant starch, 51 SNPs were associated with AM, and nine SNPs were associated with the AM/AP ratio. The Gene Ontology (GO) analysis identified 12 candidate genes at five QTLs associated with starch metabolism within the 200-kb intervals, located on chromosomes 1, 5, 6, and 9. Of these genes, Sobic.006G036500.1 encodes peptidyl-prolyl cis-trans-isomerase CYP38 responsible for hexose monophosphate shunt (HMS) and Sobic.009G071800 encodes 6-phospho-fructokinase (PFK), which is involved in the embden-meyerhof pathway (EMP). Kompetitive allele specific PCR (KASP) markers were developed to validate the GWAS results. The C allele is correlated with a high starch content, while the T allele is linked with a low level of starch content, and provides reliable haplotypes for MAS in sorghum quality improvement.     

大口黑鲈IGF-I基因内含子1、3和4序列多态性研究

根据大口黑鲈IGF-I基因的cDNA序列设计引物,克隆IGFI基因内含子核苷酸序列,采用PCR产物直接测序方法,在中国养殖群体和美国野生群体中筛选IGF-I基因内含子上的多态位点。应用RFLP、CRS—RFLP和SSCP技术建立多态位点的检测方法,同时比较分析其中4个多态位点在两个群体中基因频率分布。结果表明：（1）IGF-I基因内含子1、3和4序列长分别为1317bp、712bp和1941bp;（2）在3个内含子上共发现7个多态位点,其中在内含子1的208和1070位为G—A突变;内含子3的第40个碱基为一个“A”的插入-缺失突变,第307位为C—T突变,683位是G—A突变;内含子4上的696碱基处有一个20bp的插入-缺失突变,在1563位为G—A突变,表明大口黑鲈IGF-I基因序列上存在较多的SNPs;（3）内含子1上SNPG1070A的A等位基因能为HindIII限制性内切酶识别,采用RFLP技术分型。根据内含子1上SNPG208A侧翼序列,设计错配引物,使错配碱基和A等位基因共同形成TaqI酶切位点,建立CRS—RFLP检测方法。内含子4上SNPG1563A采用SSCP检测方法,同时对内含子4上的插入-缺失突变进行PAGE电泳分型。试验结果显示,RFLP、CRS—RFLP和SSCP三种SNP检测方法简单易行,适于在水产动物SNP标记研究中推广应用;（4）在中国养殖群体中,只有内含子1上的SNPG1070A具有多态性,其它3个多态位点只在美国野生群体中存在多态,证实中国养殖群体的遗传多样性相对较低。