opaque-2玉米近等基因系的构建与赖氨酸含量快速检测

我国高赖氨酸玉米种质资源狭窄,opaque-2(o2)突变基因能大幅提高玉米赖氨酸含量,通过分子标记辅助选择构建o2玉米近等基因系并检测其赖氨酸含量具有重要意义。其中,要解决的两个关键问题是如何准确地将不同供体的o2突变基因导入受体系和如何快速地检测导入系的赖氨酸含量。本研究利用O2基因内紧密连锁的SSR共显性标记引物phi057检测玉米供体和受体自交系的多态性,利用其特异性和共显性构建o2近等基因系;参考已有研究,改进染料结合(DBL)法,测定18组构建成功的o2近等基因系的赖氨酸含量。结果表明,不仅在不同供体(CA339和山东2548)之间存在多态性,而且在不同受体系间也存在多态性,利用phi057能够成功地将不同供体的o2突变基因导入受体系,构建o2近等基因系;改进的DBL法分析表明,不同受体系赖氨酸含量变化较大,不同背景的受体系导入o2突变基因后赖氨酸含量增加的幅度差异也较大;普通玉米自交系间赖氨酸含量为0.223%~0.368%,构建成功的不同o2近等基因系间,赖氨酸含量为0.373%~0.527%,与受体亲本相比,赖氨酸增加幅度最低为13%,最高为74%。分析表明,phi057能准确筛选导入o2突变基因的受体系,结合改进的DBL法能快速地选择赖氨酸含量高的玉米。     

不同浓度赖氨酸对猪IEC碱性氨基酸转运载体mRNA表达的影响

本研究通过建立猪肠粘膜上皮细胞（Intestinal epithelial cells, IEC）原代培养的方法,研究了赖氨酸对体外培养的猪小肠粘膜上皮细胞碱性氨基酸转运载体mRNA表达的影响。用机械刮取初生仔猪小肠粘膜加胶原酶Ⅱ消化的方法,成功地在体外培养了新生仔猪IEC;分别用浓度为0.5 mM、2 mM、8 mM赖氨酸处理细胞96 h,提取细胞总RNA,采用实时荧光定量PCR方法,检测了碱性氨基酸转运载体在不同处理细胞中的表达丰度。结果表明：高浓度的赖氨酸对CAT-1 mRNA的表达有上调作用,差异极显著(P<0.01),并且呈剂量依赖效应;不同浓度的赖氨酸对CAT-2 mRNA的表达影响不大,8 mM和2 mM赖氨酸组CAT-2 mRNA的表达丰度均有低于0.5 mM的趋势,但二者之间差异不显著(P>0.05);不同浓度的赖氨酸对y+LAT1 mRNA表达影响差异不显著(P>0.05);高浓度的赖氨酸可提高4F2hc mRNA的表达;0.5 mM和2 mM赖氨酸组4F2hc mRNA的表达丰度均低于8 mM,且差异显著(P<0.05);0.5 mM和2 mM赖氨酸组之间差异不显著(P>0.05)。     

玉米高蛋白高赖氨酸新种质的选育及相关性研究

饲养业迫切要求高蛋白质又高赖氮酸的“双高”玉米，我们用高蛋白质种源与高赖氮酸种源杂交，在其后代中分别用选株自交、回交、系内混合授粉等方法结合蛋白质、赖氨酸的测定来选择“双高”优株，并对自交后代进行有关相关性研究。现已选育出一小批蛋白质含量１５．７９％～２２．８８％，赖氨酸含量０．４６％～０．６４％的双高种质；在相关关系分析中证明了籽粒蛋白质和赖氨酸含量呈显著正相关关系；在蛋白质四种组分分析中证明０２基因同样可以调整高蛋白质中四种蛋白的比例，富含赖氨酸的谷蛋白占粗蛋白的比例大幅度增加，上升到４０％左右，而赖氨酸含量极少的胶蛋白占粗蛋白的比例由普通玉米的５０％下降到２０％左右，可以认为，通过遗传育种手段可以同步提高玉米籽粒中的蛋白质和赖氨酸含量。     

杜大长生长肥育猪赖氨酸需要的研究

为修订我国猪饲养标准,选用华南地区杜大长生长肥育猪,测定了其在20～100kg阶段的胴体指标,用无脂瘦肉生长速度预测了赖氨酸需要参数,结果表明应用实测的无脂瘦肉生长速度预测的赖氨酸需要参数与生产实际接近。用实测的无脂瘦肉生长速度建立了赖氨酸需要量的数学模型：真可消化赖氨酸需要量(mg／d)：7652．7059 115．3600m 1．1718m^2—O．0188m^3 36．0000m^0.75(m为体质量)。     

山东省近期推广小麦品种的品质分析与分类

本研究对山东省近几年推广的24个小麦品种。包括在省区试中表现较好的尚未定名的8个新品系。按照评价小麦品质的“最优协调方案”进行了粗蛋白、赖氨酸、面筋和沉淀值的测定。并利用系统聚类方法在计算机上进行了单项指标和四项综合考虑的合理分类。     

Marker‐assisted introgression of rare allele of β‐carotene hydroxylase (crtRB1) gene into elite quality protein maize inbred for combining high lysine,tryptophan and provitamin A in maize

Vitamin A deficiency in humans is a widespread health problem. Quality protein maize (QPM) is a popular food rich in lysine and tryptophan, but poor in provitamin A (proA). Here, we report the improvement of an elite QPM inbred, HKI1128Q for proA using marker‐assisted introgression of crtRB1‐favourable allele. HKI1128 was one of the parental lines of three popular hybrids in India and was converted to QPM in our earlier programme. Severe segregation distortion for crtRB1 was observed in BC₁F₁, BC₂F₁ and BC₂F₂. Background selection by 100 SSRs revealed mean recovery of 91.07% recurrent parent genome varying from 88.78% to 93.88%. Across years, introgressed progenies possessed higher mean β‐carotene (BC: 9.22 µg/g), β‐cryptoxanthin (BCX: 3.05 µg/g) and provitamin A (proA: 10.75 µg/g) compared to HKI1128Q (BC: 2.26 µg/g, BCX: 2.26 µg/g and proA: 3.38 µg/g). High concentration of essential amino acids, viz. lysine (mean: 0.303%) and tryptophan (0.080%) in endosperm, was also retained. Multi‐year evaluation showed that introgressed progenies possessed similar grain yield (1,759–1,879 kg/ha) with HKI1128Q (1,778 kg/ha). Introgressed progenies with higher lysine, tryptophan and proA hold immense potential as donors and parents in developing biofortified hybrids.     

农杆菌介导的转高赖氨酸蛋白基因（sb401）水稻T4代分析

通过农杆菌介导的方法用富赖氨酸蛋白基因（sb401）转化粳稻品种日本晴,获得了独立的10个转化株系,对转化株系进行连续自交,通过筛选得到9个纯合的T4代转化株系。 Southern blotting分析发现,整合位点是随机的,并为低拷贝（1~3个）。TAI-PCR扩增得到8个T-DNA侧翼序列,并定位于日本晴的7条染色体上。蛋白质和氨基酸测定分析发现,sb401基因对各株系的蛋白质、赖氨酸和其他氨基酸组分的提高起到了一定的作用。将杂交结果与T-DNA插入位置结合分析发现,在低拷贝的情况下,表达量的差异不明显。     

Agronomic Practices Influence Maize Grain Quality

Abstract

Maize grain yields have increased dramatically over the past fifty years, and concurrently end-uses have proliferated requiring special quality characteristics. Plant breeders have developed many specialty types of maize, all of which are influenced by the agronomic practices used to produce the crop. Grain yield increases have resulted in lower protein concentration except when the yield increase resulted from nitrogen fertilizer application. Irrigation improves the biological value of protein, while higher nitrogen application rates alter the amino acid balance thereby reducing the nutritional value. Kernel breakage susceptibility and kernel density increase with higher nitrogen fertilizer application rates, and are reduced by irrigation. Extractable starch and oil concentration are largely influenced by hybrid choice, but small production practice effects have been documented. Essential mineral nutrient levels are often influenced by soil or foliar fertilizer application. The production of aflatoxin can be reduced by irrigation or other strategies to avoid water stress during grain fill. Although genetics usually exerts the largest effect on maize grain quality, agronomic practices are also important.     

Effects of different arginine/lysine level on growth performance,body composition and digestive enzyme activity of Macrobrachium rosenbergii

Given the universal presence of antagonism between arginine and lysine, determining the optimal dietary arginine/lysine level is of great importance. In this study, eight diets were formulated to contain different arginine/lysine level, followed by a 9‐week feeding trial with shrimps of 2.10 ± 0.01 cm in length and 0.38 ± 0.02 g in weight to determine the effect of dietary arginine/lysine level on growth performance, body composition and enzyme activity. The results showed that the Diet 6 (arginine/lysine level of 29.4/21.0 g/kg) group had significantly higher weight gain rate, and specific growth rate than the control group and most of the other groups (p < .05) did. Similarly, the retention efficiency of arginine and lysine in the Diet 6 group was significantly higher than the control group and most of the other groups (p < .05). Moreover, most digestive enzymes had higher activity levels in the Diet 6 group, and evident antagonism between dietary arginine and lysine was detected. In summary, dietary arginine/lysine level of 29.4/21.0 g/kg produced the best growth performance, suggesting that a better growth performance in Macrobrachium rosenbergii culture can be obtained by adjusting the dietary arginine/lysine level.     

Dietary lysine requirement of juvenile Protonibea diacanthus

An 8‐week experiment was conducted to determine the optimal dietary lysine requirement for juvenile Protonibea diacanthus. Six isonitrogenous and isolipidic diets were formulated to contain levels of 10.8 (L10.8), 18.2 (L18.2), 26.1 (L26.1), 33.9 (L33.9), 40.7 (L40.7) and 48.6 g/kg (L48.6) of diets and were fed to the juvenile Protonibea diacanthus, respectively. The results indicated that weight gain (WG), specific growth rate (SGR), protein efficiency ratio (PER) and final weight (FW) increased as the dietary lysine level increased from 10.8 to 26.1 g/kg and then decreased as the dietary lysine levels further increased (p < .05). The lowest feed conversion ratio (FCR) was found when dietary lysine level was 26.1 g/kg. Analysis of specific growth rate by two slope broken‐line model indicated that the estimated optimal dietary levels of lysine for juvenile Protonibea diacanthus was 23.06 g/kg (51.24 g/kg dietary protein).