2002年我国大豆(Glycine max)品种及种质资源的蛋白质和脂肪含量分析

对2002年全国14个主要大豆种植省(区)的大豆品种及其种质资源的蛋白质、脂肪含量进行了分析。结果表明,种质资源的蛋白质、脂肪平均含量均高于生产用品种。其中种质资源脂肪含量的变化幅度较大,显示了在未来的育种中更强的高脂肪含量的选择优势。国内种质资源蛋白质平均含量高于国外种质资源。国外种质资源脂肪含量总体上高于国内种质资源。黄淮海生态区品种蛋白质平均含量高于北方生态区品种。新育成黄淮海区域试验品种蛋白质平均含量高于目前黄淮海生态区生产用品种。同品种异地种植,其脂肪含量有明显变化。     

大豆种子发育过程中异黄酮的积累

3个大豆品种种子中的异黄酮含量随着种子的发育进程而逐渐提高,尤其在种子成熟的最后1周内,异黄酮含量迅速增加,且与发育天数是显著正相关,与水分含量呈极显著负相关。在种子积累的不同异黄酮组分中,以雨二酰基结合体的含量最多,达总量的65％左右,而游离态积累的较少。     

花生种子的发育与贮藏蛋白质的合成和积累

花生果针入土后16天(16 DAP),种子干重和鲜重开始迅速增加。整个发育阶段可分为5个时期:组织分化期(0～20 DAP)、成熟前期(21～28 DAP)、成熟中期(29～40DAP)、成熟中后期(41～62 DAP)和成熟后期(63～88DAP)。种子发芽率在成熟前期和中期迅速提高并到达最大值,而苗成活率在成熟中后期达到最大值,苗鲜重则以88 DAP种子的为最大。种子发育过程中,贮藏蛋白质的合成与积累模式与种子干重变化相似。SDS-PAGE分析表明,种子发育初期(16 DAP)子叶中已积累花生球蛋白和伴花生球蛋白I。双向凝胶电泳显示花生球蛋白各个亚基在20DAP时均已存在,伴花生球蛋白I的主要亚基在整个发育过程中其等电点有所变化,含量也逐渐增加。其他蛋白质在种子发芽力形成阶段(20～40 DAP)的变化较为显著。     

PVA和PEG预处理对大豆种子在低温吸胀过程中胚根线粒体发育和超微结构的影响

电镜观察发现,大豆种子在刚开始萌发时胚根细胞中未能见到线粒体,线粒体是在种子萌发过程中逐渐出现的,由原质体再分化发育而成。对照胚根细胞内原质体在低温吸张过程中明显膨胀,在回温后胚根细胞中原质体仍不能发育成线粒体,甚至网状膜结构破坏,呈空泡化;经聚乙烯醇(PVA)和聚乙二醇(PEG 6000)预处理的大豆种子在同样条件下线粒体能继续发育,在回温后预处理胚根细胞中线粒体发育良好,具有明显的双层膜和管状嵴的结构。这些结果表明,在低温吸胀过程中原质体能够继续再分化发育成线粒体是提高大豆种子活力和抗冷力的重要原因。     

温度对不同类型早籼稻灌浆期间直链淀粉、蛋白质积累的影响

高温条件下早籼稻籽粒直链淀粉积累速率快,持续时间短;蛋白质含量高;灌浆前期谷草转氨酶和谷丙转氨酶活性高,开花11d后迅速下降．适温条件下籽粒直链淀粉积累速率慢,持续时间长;蛋白质含量低;谷草转氨酶和谷丙转氨酶活性变化缓慢,3个不同类型品种中适合作米粉的湘早籼33号直链淀粉含量最高,饲料稻湘早籼24号蛋白质含量最高．高温有助于湘早籼33号直链淀粉含量和湘早籼24号蛋白质含量的提高。     

PVA和PEG(6000)预处理对大豆胚轴生长、蛋白质合成和ATP含量的影响

在低温吸胀阶段,经PVA(聚乙烯醇)和PEG(聚乙二醇6000)预处理的大豆胚轴蛋白质合成和ATP含量均比对照高。在萌发阶段,胚轴生长增快,蛋白质合成明显加快,ATP迅速被消耗,而对照胚轴则相反。试验结果表明,预处理大豆种子萌发和生长与其蛋白质合成、ATP水平和消耗能力有密切关系。     

渗透调节对冷敏感大豆[Glycine max(L.)Merr.]种子膜类脂组成和脂肪酸含量的影响

选用冷敏感的大豆[Gzycine max（L.）Merr．]品种“中黄22”为试材,运用薄层层析法和气相色谱法研究了渗控增强大豆种子活力过程中膜类脂的组成及变化规律,揭示生物膜类脂在抗吸胀冷害中的作用。结果表明,用33％（W／V）的PEG6000处理种子,随着渗控时间的延长,种子的发芽指数、活力指数和根干重都逐渐增加,在渗控72h时,大豆种子中不饱和磷脂如PC、PE的比例增加,各极性脂的脂肪酸组成也发生了变化,PC、PE和PI中饱和脂肪酸16：0的含量显著下降,而不饱和脂肪酸18：2明显上升,使它们的不饱和指数大大提高,并与渗控时间呈正相关。由以上结果可证明,经PEG渗控处理后,提高了冷敏感大豆膜的流动性,为防止吸胀冷害,保持膜系统的完整性和生物膜正常的代谢提供了前提条件。     

花后土壤水分状况对小麦籽粒淀粉和蛋白质积累关键调控酶活性的影响

在温室盆栽条件下,以2个不同蛋白质含量的冬小麦(Triticum aestivum L．)品种皖麦38和扬麦9为材料,研究了花后第4天开始的土壤干旱(SRWC=45％～50％)和渍水对籽粒蛋白质和淀粉积累关键调控酶活性的影响。小麦叶片和籽粒的测定结果均表明,小麦源库器官中籽粒蛋白质和淀粉积累的关键调控酶活性变化趋势在2个品种间基本一致。与对照(SRWC=75％～80％)相比,干旱和渍水均明显降低了花后旗叶中蔗糖含量和磷酸蔗糖合成酶(SPS)活性,而氨基酸含量和谷氨酰胺合成酶(GS)活性略有下降。干旱和渍水均降低了籽粒库蔗糖合成酶(SS)和结合态淀粉合成酶(GBSS)活性,可溶性淀粉合成酶(SSS)活性降低尤甚。其中干旱处理下SS的下降比渍水更为明显。与对照相比,渍水明显降低了籽粒谷丙转氨酶(GPT)和GS活性,而干旱的影响较小。相关性分析结果表明籽粒淀粉产量和含量与SPS,SSS和GBSS活性的关系比与SS活性的关系更为密切,籽粒蛋白质产量和含量与叶中GS和籽粒中GPT活性的关系比与籽粒中GS关系活性更为密切。这些结果表明小麦源库器官中调控籽粒蛋白质和淀粉积累的关键酶活性变化是花后不同水分状况影响籽粒淀粉和蛋白质特性的重要因素。     

脱落酸对发育中花生胚萌发和贮藏蛋白质合成的影响

发育中的花生胚在无激素固体培养基上高体培养时提前萌发,其发芽力随胚的成熟增加而提高。果针入土后40d胚的发芽率达100％。禹体培养过程中,外源ABA能够阻止花生胚提前萌发和促进胚的发育。胚成熟前期,较低浓度的ABA(10~(-5)mol/L)便抑制胚的萌发;而在成熟中期以后,则要求较高浓度的ABA(10~(-4)mol/L)才能抑制胚的萌发。ABA对成熟前期胚的贮藏蛋白质合成无影响,而对成熟中期至后期胚的贮藏蛋白质合成起促进作用。ABA维持花生胚贮藏蛋白质合成和积累的作用表现在转录水平上。     

温度对美洲斑潜蝇发育、存活和繁殖的影响

在恒温１５,２０,２５,３０和３５℃下,对美洲斑潜蝇在菜豆上的发育和繁殖做了研究。结果表明,在１５￣３５℃范围内,未成熟期各虫态（卵、幼虫和蛹）的发育历期随温度的升高而缩短,发育速率呈逻辑斯蒂曲线变化;卵期孵化率在２５℃中最高;各温度下幼虫期存活率最高,低温１５℃和高温３５℃对蛹的存活有显示抑制作用,１５℃时蛹的死亡率达３９．６％。成虫寿命一般在２０ｄ以内,与温度呈直线负相关;繁殖力最高可达２８１     

Development of protein bodies and accumulation of carbohydrates in a soybean (Leguminosae) shriveled seed mutant

The soybean seed mutant T311, when grown under specific environmental conditions, produces shriveled seed. This research investigated changes in development of protein bodies and accumulation of carbohydrates during seed development by comparing the mutant with P2180 seeds. The shriveled seeds contained larger protein bodies but fewer protein bodies per cell than round seeds. Protein bodies in T311 seeds included more dispersed crystals and less globoid regions than P2180 seeds. The elemental compositions of the crystals and of whole seeds in T311 were different from that in P2180 seeds. Starch breakdown was reduced with concomitant lower soluble sugar content in T311 seeds after the D11 stage (10.0-11.9 mm long seeds). The reduced starch breakdown and lowered soluble sugar content were consistent with lower a-amylase activity and earlier and greater water loss in T311 seeds. Changes in development of protein bodies and accumulation of carbohydrates were associated with the development of the shriveled seeds.     

Quantitative proteomic analyses reveal the dynamics of protein and amino acid accumulation during soybean seed development

Using high throughput tandem mass tag (TMT) based tagging technique, we identified 4172 proteins in three developmental stages: early, mid, and late seed filling. We mapped the identified proteins to metabolic pathways associated with seed filling. The elevated abundance of several kinases was observed from the early to mid-stages of seed filling, indicating that protein phosphorylation was a significant event during this period. The early to late seed filling stages were characterized by an increased abundance of proteins associated with the cell wall, oil, and vacuolar-related processes. Among the seed storage proteins, 7S (β-subunit) and 11S (Gy3, Gy4, Gy5) steadily increased in abundance during early to late stages of seed filling, whereas 2S albumin exhibited a decrease in abundance during the same period. An increased abundance of proteases, senescence-associated proteins, and oil synthesis proteins was observed from the mid to late seed filling stages. The mid to late stages of seed filling was also characterized by a lower abundance of transferases, transporters, Kunitz family trypsin, and protease inhibitors. Two enzymes associated with methionine synthesis exhibited lower abundance from early to late stages. This study unveiled several essential enzymes/proteins related to amino acid and protein synthesis and their accumulation during seed development. All data can be accessed through this link: https://massive.ucsd.edu/ProteoSAFe/dataset.jsp?task=38784ecbd0854bb3801afc0d89056f84 . (Accession MSV000087577)     

施用氮肥对油用牡丹叶片氮素吸收积累与籽粒品质的影响

利用田间栽培试验,研究0 (对照)、18、24和30 g N·m^-2 4个氮肥用量对油用牡丹“凤丹”叶片氮素吸收转运以及籽粒产量和品质的影响.结果表明: 施用氮肥处理牡丹株高、冠幅、花径和花干质量与对照相比均显著增加,其中,24和30 g·m^-2氮肥处理株高比对照分别增加14.7%和15.2%.施用氮肥提高了牡丹籽粒的相关指标,24和30 g·m^-2氮肥处理籽粒产量达到最大,分别比对照增加15.2%和15.4%.施用氮肥明显增加了叶片氮素积累量、叶片氮素转移量和籽粒氮素积累量.其中,24 g·m^-2氮肥处理叶片氮素对籽粒贡献率最大.与对照相比,施用氮肥明显提高了籽粒蛋白氮、总氨基酸,以及部分饱和脂肪酸和不饱和脂肪酸的含量.在本试验条件下,施氮量为24 g N·m^-2时,叶片积累氮素向籽粒的转移量、转移率和贡献率均达到较高水平,籽粒产量较高,并且蛋白氮、氨基酸含量和不饱和脂肪酸含量也相对较高.     

The role of light in soybean seed filling metabolism

Soybean (Glycine max) yields high levels of both protein and oil, making it one of the most versatile and important crops in the world. Light has been implicated in the physiology of developing green seeds including soybeans but its roles are not quantitatively understood. We have determined the light levels reaching growing soybean embryos under field conditions and report detailed redox and energy balance analyses for them. Direct flux measurements and labeling patterns for multiple labeling experiments including [U‐¹³C₆]‐glucose, [U‐¹³C₅]‐glutamine, the combination of [U‐¹⁴C₁₂]‐sucrose + [U‐¹⁴C₆]‐glucose + [U‐¹⁴C₅]‐glutamine + [U‐¹⁴C₄]‐asparagine, or ¹⁴CO₂ labeling were performed at different light levels to give further insight into green embryo metabolism during seed filling and to develop and validate a flux map. Labeling patterns (protein amino acids, triacylglycerol fatty acids, starch, cell wall, protein glycan monomers, organic acids), uptake fluxes (glutamine, asparagine, sucrose, glucose), fluxes to biomass (protein amino acids, oil), and respiratory fluxes (CO₂, O₂) were established by a combination of gas chromatography‐mass spectrometry, ¹³C‐ and ¹H‐NMR, scintillation counting, HPLC, gas chromatography‐flame ionization detection, C:N and amino acid analyses, and infrared gas analysis, yielding over 750 measurements of metabolism. Our results show: (i) that developing soybeans receive low but significant light levels that influence growth and metabolism; (ii) a role for light in generating ATP but not net reductant during seed filling; (iii) that flux through Rubisco contributes to carbon conversion efficiency through generation of 3‐phosphoglycerate; and (iv) a larger contribution of amino acid carbon to fatty acid synthesis than in other oilseeds analyzed to date.     

Natural variation of GmFATA1B regulates seed oil content and composition in soybean

Soybean (Glycine max) produces seeds that are rich in unsaturated fatty acids and is an important oilseed crop worldwide. Seed oil content and composition largely determine the economic value of soybean. Due to natural genetic variation, seed oil content varies substantially across soybean cultivars. Although much progress has been made in elucidating the genetic trajectory underlying fatty acid metabolism and oil biosynthesis in plants, the causal genes for many quantitative trait loci (QTLs) regulating seed oil content in soybean remain to be revealed. In this study, we identified GmFATA1B as the gene underlying a QTL that regulates seed oil content and composition, as well as seed size in soybean. Nine extra amino acids in the conserved region of GmFATA1B impair its function as a fatty acyl–acyl carrier protein thioesterase, thereby affecting seed oil content and composition. Heterogeneously overexpressing the functional GmFATA1B allele in Arabidopsis thaliana increased both the total oil content and the oleic acid and linoleic acid contents of seeds. Our findings uncover a previously unknown locus underlying variation in seed oil content in soybean and lay the foundation for improving seed oil content and composition in soybean.     

Pattern analysis suggests that phosphoenolpyruvate carboxylase in maturing soybean seeds promotes the accumulation of protein

ABSTRACT

The protein and oil contents in soybean seeds are major factors in seed quality. Seed proteins and oils are synthesized from sucrose and nitrogenous compounds transported into maturing seeds. In this study, we compared changes in the activity of phosphoenolpyruvate carboxylase (PEPC) and the accumulation profiles of protein and oil in maturing seeds of two soybean cultivars, which exhibit different protein and oil contents in seeds, to determine the interrelationships of them. A principal component analysis indicated a concordance of seed PEPC activity with the protein content, but did not with the oil content. PEPC activity per seed was highest in the late maturation stage, when the physiological status of the vegetative organs drastically changed. The high-protein cultivar had higher PEPC activity compared to the low-protein cultivar. These results highlight the biological role of PEPC in the synthesis of protein, therefore it was implied that PEPC could be a biomarker in soybean breeding.

Abbreviations: ANOVA: analysis of variance; DS: developmental stage; DW: dry weight; FW: fresh weight; NIR: near infrared; PEP(C): phosphoenolpyruvate (carboxylase); PC(A): principal component (analysis); S.E.: standard error; WC: water content.     

Effects of defoliation on seed protein concentration in normal and high protein lines of soybean

Two high (NC106, NC111) and two normal (NC103, NC107) seed protein concentration lines, derived from two different recurrent selection populations of soybean (Glycine max L. Merr.) were subjected to partial defoliation at beginning seed fill (R5) under outdoor pot culture and field conditions. The aim of this study was to test the hypothesis that capacity to store N in vegetative organs and/or to mobilize that N to reproductive organs is associated with the high seed protein concentration trait. Symbiotic N₂ fixation was the sole source of N in the pot experiment and the major source of N (met > 50% of the N requirement) in the low N soil used in the field experiment. Seed protein concentration and seed yield at maturity in both experiments and N accumulation and mobilization between R5 and maturity in the pot experiment were measured. The four genotypes did not differ significantly with respect to the amount of N accumulated before beginning seed fill (R5). Removal of up to two leaflets per trifoliolate leaf at R5 significantly decreased the seed protein concentration of NC107/111 but had no effect on this trait in NC103/106. Defoliation treatments significantly decreased seed yield, whole plant N accumulation (N₂-fixation) during reproductive growth and vegetative N mobilization of all genotypes. Differences in harvest indices between the high and low protein lines accounted for approximately 35% of the differences in protein concentration. The two normal protein lines mobilized more vegetative N to the seed (average. 5.26 g plant^–1) than the two high protein lines (average. 4.28 g plant^–1). The two high seed protein lines (NC106, NC111) exhibited significantly different relative dependencies of reproductive N accumulation on vegetative N mobilization, 45% vs. 29%, in the control treatment. Whereas, NC103 with normal and NC106 with high seed protein concentration exhibited similar relative dependencies of reproductive N accumulation on vegetative N mobilization, (47% vs. 45%). Collectively, these results indicate that N stored in shoot organs before R5 and greater absolute and relative contribution of vegetative N mobilization to the reproductive N requirement are not responsible for the high seed protein concentration trait.Abbreviations DAT days after transplanting - R5 fifth reproductive stage according to Fehr and Caviness, 1977 Mention of trademark or proprietary product does not constitute a guarantee or warranty of the product by the United States Department of Agriculture and does not imply its approval to the exclusion of other products that may also be suitable.     

Protein and oil concentration of soybean seed cultured in vitro using nutrient solutions of differing glutamine concentration

Oil and protein are the most valuable components of soybean seed. Evidence indicates that growth and composition of soybean seed are controlled by supplies of carbon and nitrogen provided by the maternal plant to the seed, but it is difficult experimentally to control and quantify the precise amount of carbon and nitrogen provided to the seed by the whole plant. To examine whether oil and protein concentrations are affected by the supply of nitrogen to the seed, immature soybean seeds (Glycine max cv. Williams 82) were grown in vitro in nutrient solutions containing 20, 40, 60 or 80 mM of glutamine. The seeds were incubated in Erlenmeyer flasks for 8 days at 25°C. The rate of dry matter accumulation changed from 7.2 to 8.3 mg seed⁻¹ day⁻¹ as the glutamine concentration increased from 20 to 80 mM but the differences were not significant (P 0.05). Seed protein concentration increased as glutamine concentration increased from 294 mg g⁻¹ at 20 mM glutamine to as high as 445 mg g⁻¹ at 80 mM glutamine. Typical in vivo protein concentration of mature soybean seeds is about 400 mg g⁻¹. Oil and protein concentrations were negatively correlated (r²= 0.44), which indicates that oil and protein synthesis are interrelated. Protein synthesis was favoured over oil synthesis when nitrogen became more abundant. The seeds used in this study clearly demonstrated a capacity to respond to nitrogen availability with changes in seed protein concentration.     

Synthesis and degradation of the major allergens in developing and germinating soybean seed

Gly m Bd 28K,Gly m Bd 30K and Gly m Bd 60K are the major soybean(Glycine max(L.)Merr.)allergens limiting the consumption of a good protein source for sensitive individuals.However,little is known about their temporal-spatial expression during seed development and upon germination.The present data shows that soy allergens accumulated in both the embryonic axes and cotyledon,but expression patterns differed depending on the specific allergen.Allergens accumulated sooner and to a greater level in cotyledons than in embryonic axes.Gly m Bd 28 began at 14 d after flowering,7 to 14 d earlier than Gly m Bd 30K and Gly m Bd 60K.Comparatively,their degradation was faster and more profound in embryonic axes than in cotyledons.Gly m Bd 60K began to decline at 36 h after imbibition and remained detectable up to 108 h in cotyledons.In contrast,the Glym Bd 60K protein was reduced at 24 h,and eventually disappeared at 96 h.In cotyledons Gly m Bd 28K first declined at 24 h,then increased from 36 h to 48 h,followed by its large reduction at 72 h after seed germination.These findings provide useful information on soy allergen biosynthesis and will help move forward towards developing a hypoallergenic soybean for safer food.     

西伯利亚白刺籽油的抗疲劳活性

目的考察西伯利亚白刺籽油的抗疲劳活性,为开发利用白刺提供依据。方法以西伯利亚白刺籽油为供试品,以ICR小鼠为受试动物,分为空白对照组、阳性对照组和样品组低、中、高3个剂量进行小鼠负重游泳实验,观察样品对小鼠抗疲劳的影响。结果西伯利亚白刺籽油高剂量组小鼠负重游泳时间显著延长（P〈0.05）,运动后血乳酸含量比空白对照组小鼠减少40%,同时肝糖元含量显著提高。结论西伯利亚白刺籽油在提高抗疲劳性方面有一定的功效。