我国江浙地区粳稻品种间的植酸与蛋白质组分差异及其相关性

选用我国江浙地区近期育成的晚粳稻品种（系）29个,对它们在浙江杭州相同栽培条件下籽粒中的植酸、蛋白质含量与其组分差异及其相关性进行了分析。供试29个品种(系)植酸含量的平均值为0.868%,变化幅度为0.699%~1034%,其中秀水系列品种的植酸含量一般较低,而武育粳系列和淮字系列品种（系）的植酸含量则相对较高;供试29个品种材料总蛋白质含量的平均值为8.722%,其含量大致呈正态分布。在4种蛋白质组分中,谷蛋白、球蛋白和清蛋白在品种间的变异系数较大,而醇溶蛋白则相对较小,且在分布频率上也表现出品种类型间的差异特征;除谷蛋白与总蛋白质之间存在着极显著的正相关外,供试29个品种(系)的植酸含量与蛋白质含量及4种蛋白质组分含量间并无密切的关系。     

Positional variations in phytic acid and protein content within a panicle of japonica rice

Six japonica rice cultivars differing in phytic acid and protein content were used to investigate the effect of grain position in the panicle on grain weight, phytic acid and protein content. There were significant differences in grain weight, albumin, globulin, prolamin, glutelin, and total protein content among the grains within a panicle for all six cultivars, but no differences were found for phytic acid content. An effect of grain position between primary rachises on grain weight and protein content was clearly detected, while that among top, middle and bottom rachis showed a greater change, varying with the cultivar. Albumin and globulin contents tended to be lower in the grains on the primary rachis than in those on the secondary rachis, and glutelin and total protein contents showed the opposite trend in the three cultivars with lower protein content. There was no significant correlation between phytic acid and protein content in the different grain positions within a panicle. The accumulation of phytic acid and protein during grain development appears to be independent.     

稻米蛋白质组分及总蛋白质含量与淀粉RVA谱特征值的关系

 利用低谷蛋白含量（low glutelin content, LGC）粳稻品种W1721与大面积应用的杂交粳稻恢复系轮回422构建了重组自交系群体。根据种子总蛋白质的SDS PAGE带型,推断各单株LGC位点的基因型。在排除脂肪的影响下,比较了基因型LGCLGC（谷蛋白含量较醇溶蛋白低）与基因型lgclgc（谷蛋白含量较醇溶蛋白高）在RVA谱特征值上的差异,以及同一基因型内总蛋白质含量与淀粉RVA谱特征值的关系。结果表明基因型LGCLGC的崩解值和回复值的平均值分别显著和极显著大于基因型lgclgc;LGC对崩解值和回复值的相对加性效应分别为3.91%和4.45%,贡献率分别为4.81%和12.81%。LGCLGC型的稻米,总蛋白质含量与崩解值和消减值分别存在显著的负线性回归和正线性回归关系,贡献率分别为9.86%和11.48%;lgclgc型的稻米,总蛋白质含量与消减值和回复值都存在极显著的负线性回归关系,贡献率分别为1341%和27.88%。结合前人研究认为,稻米食味品质受谷蛋白相对于醇溶蛋白的含量以及总蛋白质含量的影响;而总蛋白质含量对食味品质的影响因谷蛋白相对于醇溶蛋白的含量的不同而异。因此,育种工作中应当把谷蛋白相对于醇溶蛋白的含量以及总蛋白质含量同时作为选择指标。     

水稻籽粒不同部位植酸含量及其与稻米品质的相关性

 选用13个不同类型的水稻品种,对其籽粒不同部位植酸含量品种间差异及与稻米主要品质性状间的相关性进行了分析。结果表明,供试13个水稻品种米糠、糙米、颖壳、精米的植酸含量平均值分别为48.51、9.77、1.40和0.91 mg/g,而且不同水稻品种间籽粒不同部位的植酸含量存在极显著差异。在籽粒中,米糠中植酸积累最多,其次是精米,颖壳最少。米糠中植酸含量与糙米和颖壳中植酸含量呈极显著正相关。糙米和米糠中植酸含量与稻米蛋白质含量均呈显著的负相关,与千粒重均呈显著正相关。     

QTL mapping for crude protein and protein fraction contents in rice (Oryza sativa L.)

Improvement of rice storage proteins is important in rice breeding for high nutritional quality. Seventy-one recombinant inbred lines (RIL) derived from a cross between japonica variety Asominori and indica variety IR24 were used to study the inheritance of crude protein and protein fraction contents in rice. A total of 16 QTL were identified and mapped on eight chromosomes. Several QTL affecting contents of different protein fractions were mapped in the same chromosomal region. In particular, two QTL with a significant contribution were identified to simultaneously affect prolamin and glutelin contents. One QTL denoted as qCP-12 affecting crude protein content (CP) was located in the same region as QTL qGLT-12 affecting glutelin content, in agreement with the positive correlation between glutelin level and protein content. QTL with larger genetic effects were further confirmed using two sets of chromosome segment substitution lines (CSSL), where Asominori and IR24 were used as the recurrent parents. By QTL comparative analysis, two QTL for CP, three for globulin content and one for prolamin content were located in the vicinity of CP QTL previously identified in polished rice. Application of these results in rice breeding is discussed.     

水稻种子蛋白质的组成和积累形态对稻米品质的影响

设置水培试验对‘16-17’（谷蛋白前体聚合物明显,含量高）和‘16-20’（谷蛋白前体聚合物不明显,含量低）分别进行CK（氮2800 μmol/L,硫80 μmol/L）、N（氮3640 μmol/L,硫80 μmol/L）、S（氮2800 μmol/L,硫260 μmol/L）、N+S（氮3640 μmol/L,硫260 μmol/L）处理,以此改变稻米品质和稻米贮藏蛋白质的组成与含量,从而对稻米蛋白质各组分及相关品质性状进行分析。结果表明：（1）在N、S和N+S的不同处理下均增加了粗蛋白的含量,使‘16-20’的稻米粗蛋白百分含量分别增加2.35%、2.67%和2.73%;（2）在2个品种的各组分蛋白含量变化中,‘16-17’的醇溶蛋白和谷蛋白的百分含量在N、S和N+S下较对照分别增加了9.75%、4.21%和4.67%,3.92%、7.38%和12.92%。‘16-20’的谷蛋白百分含量在S和N+S处理下增加了17.72%和29.38%,醇溶蛋白百分含量在N+S下增加了3.73%,而球蛋白百分含量在N、S和N+S三个处理下降低了2.86%、3.78%和3.69%,清蛋白在N、S处理下显著降低;（3）谷蛋白的2个亚基的相对含量和57 kDa谷蛋白前体及57 kDa以上谷蛋白聚合体的积累在‘16-20’品种中明显增加。（4）稻米品质中,N、S配施对2个参试品种的碾磨品质性状的影响不明显,但有利于2个品种稻米外观品质和食味品质的改善,并使‘16-20’的营养品质显著增加,另外,‘16-20’的千粒重在S和N+S处理下显著增加。故不同N、S配施处理下,不同品种水稻贮藏蛋白质的组成和积累形态差异很大,并改变了含巯基蛋白质组分的积累量和积累形态。尤其是在富S条件下,蛋白质积累过程中形成较多的巯基及二硫键,并通过二硫键来改变含巯基的蛋白质亚基之间的聚合程度,形成不同分子量的谷蛋白聚合体,使蛋白质的积累形态发生明显改变,进而使稻米相关品质性状发生改变。     

Effect of nitrogen fertilizer application on grain phytic acid and protein concentrations in japonica rice and its variations with genotypes

Thirty-one japonica cultivars with contrasting agronomic traits were used and 5 nitrogen (N) fertilizer treatments of level and timing were performed to determine the effects of N application on grain phytic acid and proteins including albumin, globulin, prolamin, and glutelin. Variance analysis showed a large effect of N and a smaller effect of cultivar on grain phytic acid and proteins. With increased N level, grain phytic acid concentrations progressively reduced whereas concentrations of the 4 proteins and ratios of glutelin to total protein increased, indicating that N level has a beneficial effect on rice nutritional quality. In addition, substantial genotypic differences in response of grain phytic acid and proteins to N treatments were detected. Some cultivars such as Xiushui09, Zhendao10, and Yanjing5 exhibited more stability of grain proteins under contrasting N treatments, and the significance of this stability is discussed in regard to its use in rice quality improvement.     

醋制对黑豆蛋白含量及多肽ACE抑制活性的影响

为考察醋制对黑豆蛋白含量及多肽ACE抑制活性的影响,为对黑豆醋浸过程中蛋白组分及含量变化进行分析,并对相应ACE抑制肽活性进行测定。黑豆经醋浸不同时间后,采用顺序抽提法提取清蛋白、球蛋白、醇溶蛋白和谷蛋白,并采用凯氏定氮法及SDS-PAGE电泳进行定量和定性分析;各蛋白组分经胃蛋白酶水解,经超滤(截留分子量3 k D)后收集滤液,获得多肽组分,RP-HPLC法进行ACE抑制活性评价。结果表明:经醋浸14 d后,黑豆总蛋白含量变化幅度小于±0.5%;清蛋白含量由55.13%(0 d)降低至9.61%(14 d);球蛋白由7.04%(0 d)增加到20.34%(14 d);醇溶蛋白由0.81%(0 d)增加到1.72(14 d);谷蛋白由21.11%(0 d)增加到59.45%(14 d),含量最高。醋浸处理降低了清蛋白源多肽的ACE抑制活性,但提高了球蛋白、醇溶蛋白及谷蛋白多肽的ACE抑制活性,其中,谷蛋白多肽抑制率由18.18%(0 d)增加至37.58%(14 d),抑制活性升高。醋浸可改变黑豆各蛋白组分的含量和对应多肽的ACE抑制活性,其中,谷蛋白含量及其多肽ACE抑制活性均有增加。研究结果表明可进一步采用分离纯化技术从谷蛋白多肽中获得高活性降压肽。     

杂交饲料稻组合的筛选初步研究

１９９６－１９９７年进行了饲料稻品种的筛选试验,初选出４个高产高蛋白杂交饲料稻组合共近期生产上主推应用。研究发现,杂交组合的蛋白质含量受恢复系的影响较大,产量与蛋白质含量,蛋白质含量与出糙率之间均不存在显著的直线和相关关系。     

Analysis of Storage Protein Distribution in Rice Grain of Seed-Protein Mutant Cultivars by Immunofluorescence Microscopy

Abstract

Localization of storage proteins in rice grains of seed-protein mutant cultivars, low-glutelin cultivars LGC-1, LGCsoft and a 26-kDa-globulin-deficient low-glutelin cultivar LGC-Katsu, was examined by immunofluorescence microscopy using fluorescence-labeled antibodies of 13 kDa prolamin and 23 kDa glutelin. Abundant 13 kDa prolamin and 23 kDa glutelin was observed in the outer regions of rice grains. Image analysis revealed that peaks of fluorescence intensity of both proteins were located at 2–7% of the width or longitudinal length of brown rice distant from the outer surface of brown rice (RDOS) on the dorsal, ventral, and apical sides of brown rice. In these seed-protein mutant cultivars and a normal-type cultivar Nihonmasari, the foot of the distribution peaks of both proteins were located at 20–30% RDOS on the ventral and apical side. In contrast, on the dorsal side of rice grain, varietal differences of RDOS at the foot of the distribution peak of both proteins varied with the cultivar. The RDOS was 20 –40%; and Nihonmasari >LGC-1≒LGCsoft >LGC-Katsu. Although the quantity of 13 kDa prolamin and 23 kDa glutelin greatly varied with the cultivar in the outer region of rice grain, both proteins were distributed uniformly at low levels in the inner region in all cultivars. Furthermore, SDS-PAGE analysis showed that a larger quantity of 13 kDa prolamin localized on the ventral than the dorsal side of rice grains in seed-protein mutant cultivars, especially in LGC-Katsu.     

两个环境下糙米和精米蛋白质含量的QTL分析

 利用两个直链淀粉含量相似的水稻品种93 11和日本晴为亲本,采取单粒传法创建由190个家系组成的重组自交系群体,并构建了包含202个SSR、CAP和STS标记的遗传连锁图谱。采用复合区间作图法,在两个不同生态环境下(三亚、合肥)对糙米蛋白质含量和精米蛋白质含量的QTL进行了定位分析,共定位到9个QTL。其中,qBRPC1.3在两个环境中均被检测到,稳定性较高,其贡献率达9.4%;位于第1染色体的RM578-RM10596是多效性区间,同时控制糙米蛋白质含量和精米蛋白质含量,贡献率分别为6.4%和5.7%。     

几个早籼新品种（组合）的饲用前景分析

通过５个不同生态点的品比试验，分析了５个早籼新品种（组合）的丰产性、稳产性和生育期，结合田间抗性以及室内测定的出糙率、糙米粗蛋白质和氨基酸含量，提出近期内适于作饲料稻开发的早籼良种为威优５６。     

南方和黄淮海地区大豆地方品种贮存蛋白组分含量遗传变异的初步分析

我国南方和黄淮海地区的７２个大豆地方品种的贮存蛋白组分含量分析结果表明，大豆种子贮存蛋白各组分平均含量为球蛋白．２４．３９、清蛋白５．７７、醇溶蛋白１．７４、谷蛋白５．２７（ｇ／１００ｇ），其遗传变异丰富。南方与黄淮海地区品种间的４种贮存蛋白组分含量平均数及方差均无显著差异。     

大豆种子贮存蛋白组成及其相关分析

1988年选用14个,1989年选用15个蛋白质含量差异较大的主要为江淮地区的大豆品种,进行随机区组试验。大豆种子中球蛋白、清蛋白、醇溶谷蛋白以及谷蛋白的品种间平均分别为28.56％、7.50％、1.81％和5.71％,品种间变幅分别为6.08％、10.25％、1.03％和6.51％。清蛋白含量与全蛋白含量。谷蛋白含量与全蛋白含量之间显著正相关,清蛋白含量对种子全蛋白含量品种间差异的相对贡献最大,而球蛋白含量对品种间全蛋白含量差异的相对贡献最小。球蛋白含量与株高、分枝数和单株粒重存在显著的正相关;但株高和清蛋白含量、单株荚数及单株粒数和谷蛋白含量有显著负相关。     

Effects of Temperature on the Digestible Protein Content of Grains during Ripening in a Seed-protein Mutant Rice Cultivar LGCsoft

Abstract

The effects of temperature during the ripening period on digestible protein contents of the rice grains of a seed-protein mutant rice cultivar LGCsoft were examined. The plants were grown under a natural condition until the booting stage, and then in temperature-controlled greenhouses set at 24.0ºC, 28.0ºC, and 30.6ºC (mean temperature). The protein compositions and the protein contents of the rice grains were analyzed quantitatively. The protein compositions in the LGCsoft grains varied with the temperature condition. The ratio of the digestible to total protein was higher in high-temperature conditions, and that of difficult-to-digest proteins, especially 13 kDa prolamin was lower in high-temperature conditions. The protein compositions in a normal-type cultivar Nihonmasari, which was the original cultivar of LGCsoft also varied with the temperature. However, the effect of temperature on the ratio of the digestible to total protein was larger in LGCsoft than in Nihonmasari. The ratios of the digestible protein in the grains under 24.0ºC and 30.6ºC conditions were 74.3% and 81.3%, respectively, in Nihonmasari. On the other hand, they were 52.0% and 63.1%, respectively, in LGCsoft. In LGCsoft, the total protein content of grains was 70.6-72.5 mg g-¹, and it was affected only slightly by temperature during the ripening period. Therefore, the digestible protein content of grains under 24.0ºC and 30.6ºC conditions was 36.7 mg g-¹ and 45.7 mg g-¹, respectively, in LGC soft. It was clarified that the digestible protein content was higher at elevated temperatures because of the increased ratio of digestible to total protein.     

Distribution of proteins and amino acids in milled and brown rice as affected by nitrogen fertilization and genotype

A field experiment involving six japonica rice cultivars with contrasting agronomic traits and seven nitrogen (N) fertilizer treatments was performed to determine the effects of N and genotype on distribution of four kinds of grain proteins and amino acids in milled and brown rice. For brown and milled rice, albumin and globulin were controlled more by genotype than N treatments, whereas prolamin and glutelin were largely determined by N. Substantial genotypic differences in response of milled/brown (M/B) ratios of proteins to N treatments were detected. In comparison with large panicle cultivars, small panicle cultivars such as Wuyujing3 had the lower ratio and exhibited more stability under contrasting N treatments. N had significant influence on amino acid composition of brown and milled rice, with contents of the 17 amino acids measured increasing with elevated N rate. However, cysteine and methionine in brown rice and lysine and methionine in milled rice were not significantly affected by N. In addition, N had little effect on ratios of M/B for most of the amino acids.     

Characterization and In Vitro digestibility of rice protein prepared by enzyme-assisted microfluidization: Comparison to alkaline extraction

Microfluidization followed by density-based separation was employed to extract protein from broken rice by disrupting protein-starch agglomerates. Follow-up enzyme treatments (amylase and glucoamylase) were performed to further improve the purity of the protein-rich fraction. High protein recovery (81.87%) and purity (87.89%) were obtained. The protein composition, solubility, structural properties, and in vitro digestibility of rice proteins prepared by enzyme-assisted microfluidization (EM-RP) and alkaline extraction (AE-RP) were compared. EM-RP was mainly composed of glutelin, which had low solubility and native structure. By contrast, large quantities of prolamin and globulin appeared in the AE-RP except glutelin, leading to the richness of glutamic acid/glutamine, leucine, aromatic and charged amino acids in the AE-RP. Compared to AE-RP, EM-RP showed higher digestibility due to the richness of glutelin (an easy-to-digest protein), as evidenced by higher nitrogen release during pepsin-trypsin digestion. The presence of prolamin (an indigestible protein) in AE-RP decreased protein digestibility although alkaline extraction improved its hydrolysis. These results suggest that enzyme-assisted microfluidization could be an effective technique to non-destructively and selectively extract rice glutelin.     

High Temperature During Rice Grain Filling Enhances Aspartate Metabolism in Grains and Results in Accumulation of Aspartate-Family Amino Acids and Protein Components

Global warming causes the exacerbation of rice growing environment, which seriously affects rice growth and reproduction, and finally results in the decrease of rice yield and quality. We investigated the activities of aspartate metabolism enzymes in grains, and the contents of Aspartate-family amino acids and protein components to further understand the effects of high temperature (HT) on rice nutritional quality during rice grain filling. Under HT, the average activities of aspartate aminotransferase (AAT) and aspartokinase (AK) in grains significantly increased, the amino acid contents of aspartate (Asp), lysine (Lys), threonine (Thr), methionine (Met) and isoleucine (Ile) and the protein contents of albumin, globulin, prolamin and glutelin also significantly increased. The results indicated that HT enhanced Asp metabolism during rice grain filling and the enhancement of Asp metabolism might play an important role in the increase of Asp-family amino acids and protein components in grains. In case of the partial appraisal of the change of Asp-family amino acids and protein components under HT, we introduced eight indicators (amino acid or protein content, ratio of amino acid or protein, amino acid or protein content per grain and amino acid or protein content per panicle) to estimate the effects of HT. It is suggested that HT during rice grain filling was benefit for the accumulation of Asp-family amino acids and protein components. Combined with the improvement of Asp-family amino acid ratio in grains under HT, it is suggested that HT during grain filling may improve the rice nutritional quality. However, the yields of parts of Asp-family amino acids and protein components were decreased under HT during rice grain filling.     

Seed protein fractions and amino acid composition in gram (Cicer arietinum)

Six chickpea strains were analysed for their protein content and various protein fractions. The protein content ranged from 20.9–25.27%. Albumin, globulin, prolamin and glutelin contents ranged from 8.39–12.31%; 53.44–60.29%; 3.12–6.89% and 19.38–24.40% respectively. Salt soluble proteins (albumin + globulin) and globulins resolved into 19–23 bands whereas albumin proteins resolved into 30–34 bands. The molecular weights of various polypeptides ranged from 10–91 kD. Amino acid analysis of total proteins revealed that glutamic acid was present in maximum concentration followed by aspartic acid and arginine. Just like other pulse proteins, chick pea proteins were also found deficient in sulphur containing amino acids.     

水稻籽粒蛋白质含量选择对杂种早代蒸煮食味品质的影响

选用品质和产量性状不同的4个粳稻品种配制2个杂交组合,研究籽粒蛋白质含量选择对水稻杂种早代蒸煮食味品质的影响。各品质性状变异系数的变化范围在9.73%~94.99%;直链淀粉含量与蛋白质含量间存在明显的动态关系;稻米味度值先是随着蛋白质含量的下降而增加,当蛋白质含量下降到某一临界值后,随蛋白质含量下降而下降,蛋白质与直链淀粉含量适中的后代株系味度值较高;在蛋白质含量水平较高时,稻米RVA谱特性随着蛋白质含量的下降有所改善,但蛋白质含量下降到一定程度后则又会导致RVA谱特性变劣;蛋白质含量过高或过低都会导致稻米蒸煮食味品质变劣,协调蛋白质与直链淀粉含量的平衡关系是提高稻米蒸煮食味品质的关键。