Correlation between Protein Body Formation and Storage Protein Accumulation in Soybean Cotyledons

At 20 days after flowering (DAF), the 7S α' and α subunits began to accumulate. At 25 DAF, the 7Sβ, l1SA and llSB subunits appeared. Five days later, the 11SA-4 subunit was present During the period of 25–55DAF, the storage protein content continued to increase. From 55 to 63 DAF, there was a decrease in the synthetic rate of the storage proteins. Comparing these results with the two paths of protein body formation reported previously, we draw the conclusion that the protein bodies developed from vacuoles contained not only the 7S bm also the lis proteins in soybean cotyledon cells.     

不同成熟度花生胚萌发时子叶中贮藏蛋白质的降解

花生(Arachis hypogaea L.)汕油71果针入土20d(20 DAP)的种子剥去种皮后,10％的胚可以萌发,至40 DAP发芽率达98％。不同发育时期的花生胚萌发 10d后子叶盐溶蛋白质和花生球蛋白降解表明,20和32 DAP胚萌发后,子叶中这些蛋白质只有部分降解。随着胚成熟度增加,子叶中降解这些蛋白质的能力不断提高。20～40 DAP胚萌发4d时,子叶的BAPAase和GHE活性较低。50～80DAP胚萌发 4d,子叶中上述两种酶均显示较高的活性。     

Ultrastructure and Lipase Activity of Cotyledon cell During Pod Development in Peanut

A series of significant changes of the ultrastructure and lipase activity of cotyledon cell were found in peanut (Arachis hypogaea) during pod development. In he initial stage of cotyledon development there were many plastids which kept producing starch grain and there were low lipase activity and very few lipid and protein bodies in the cell. In the middle stage of cotyledon development, a great number of larger lipid bodies were seen in the cell and a lot of protein bodies formed in the vacuoles and continued to increase in size. Lipase activity increased in the cytoplasm, endoplasmic reticulum, protein bodies, plasmalemma and intercellular space. In the later stage of cotyledon development, the lipid bodies did not increase in number but became slightly larger. The protein bodies continued to increase both in number and in size. Lipase acttvity was even hegher in the cytoplasm. In the final stage the protein bodies became irregular in shape and some of them tended to disintegrate with their content entered into the space around the lipid bodies. The lipase activity in the cell declined. The results indicated that the lipid body originated in the cytoplasm and the protein body originated in the vacuole; that the accumulation of oil and protein in peanut cotyledon resulted from the formation and development of lipid and protein bodies in the cell, and that the changes of plasmid and lipase activity in the cell played a role in the development of lipid body during the development of cotyledon.     

重组蛋白包涵体的复性研究

重组蛋白在大肠杆菌中的高表达往往形成不可溶、无生物活性的包涵体,需经过变性溶解后,在适当条件下复性形成天然的构象,才可恢复其生物活性．变复性实验是建立在对蛋白质体外折叠机制的了解的基础上．根据近年来对蛋白质折叠机制的认识和重组蛋白包涵体在复性方面的主要进展,论述以下3个方面的内容：1)蛋白质在细胞内的折叠机制;2)蛋白质体外折叠机制;3)蛋白质复性的策略和方法．     

SSR和STS标记在花生栽培品种鉴定中的应用研究

采用10份花生品种材料,对通过数据库查询设计合成的SSR引物和其他作者发表的SSR引物及STS引物进行了评估,并基于品种间简单匹配系数做了聚类分析。获得62个能产生多态性片段的SSR和STS引物对,总共获得427条带,其中291条（68．1％）为多态性带。平均每对引物产生6．88条带,4．69条为多态性带;多态性条带比率为16.7％～100％,PIC值为0．254～0．952,平均为0．760。9对SSR／STS引物,即Lec-1、Ah4-26、Ah4-4、SsS14、SHPAL-1、PG71、PG43、PM36、PG22,对所采用的10份花生品种区分率达到10096。说明SSR和STS标记应用于花生品种鉴定有效。     

去胚轴对花生子叶肽链内切酶和贮藏蛋白质降解的影响

从离体子叶与连体子叶在水中培养一段时间后的比较,看到它们之间在肽链内切酶活性和盐溶蛋白及花生球蛋白降解上的差异并不大,这表明去除胚轴对子叶肽链内切酶活性和贮藏蛋白降解的影响很轻微。亚胺环己酮(蛋白质合成抑制剂)不能完全抑制离体子叶肽链内切酶活性的提高,子叶的大部分大分子贮藏蛋白同样被降解。这表明,在花生种子萌发过程中降解大部分贮藏蛋白的子叶肽链内切酶并非全部是在种子萌发时新合成的,子叶贮藏蛋白降解和肽链内切酶活性基本不受胚轴调控,子叶与胚轴之间在调控关系上可能是一种新的调节类型。     

Ultrastructure of Protein Bodies and Plastids in Cotyledon of Nelumbo nucifera Gaertn.at Seed Germination

The present article deals mainly with the formation and dissolution of protein bodies and development of plastids in cotyledon cells of Nelumbo nucifera during seed germination. Electron microscopic studies reveal that protein bodies are formed after imbibition of the cotyledons before germination. They are produced through accumulation of protein material in small vacuoles delivered from the exudates of endoplasmic reticulum or by fragmentation of endoplasmic reticulum itself. In the period of germination, most of the material in the protein bodies dissolute and they coalesce with each other forming large vacuoles. The protein residue of the vacuoles condenses into small blocks with high electron density adhering to the tonoplast or freely floating in the vacuole. Thus, it suggests that the protein bodies of the germinating N. nucifera cotyledons are originated from vacuoles formed by endoplasmic reticulum. Part of the plastids found in cotyledonous cells of mature N. nucifera seeds exists as proplastids. They develop continuously after imbibition of the cotyledons. During the period of seed germination, many concentric lamellae are developed along the plastid membrane on which they later coalesce with the neighboring concentric lameUae forming loosely organized prolamellar bodies which condense into paracrystalline lattices. No ribosomes are present in the inter spaces of paracrystatline lattice. One to several prolamellar bodies can be developed in one plastid.     

Phenylmethylsulphonyl fluoride Inhibits the Formation of Jasmonate-Induced Proteins in Cotyledons of Cucurbita pepo (zucchini)

Phenylmethylsulphonyl fluoride (PMSF), a well known inhibitor of both thiol- and serine-type proteases, in aqueous solutions either alone or with the plant growth regulators, methyl ester of jasmonic acid (MeJA) and N⁶-benzyl-aminopurine (BAP), significantly inhibited the growth of excised Cucurbita pepo L. (zucchini) cotyledons. SDS-PAGE analysis of the protein profiles showed that PMSF suppressed the gradual decline of the main 20 – 25 kDa polypeptide group and the low molecular mass polypeptides (below 15 kDa) while leupeptine was not able to affect the electrophoretic pattern of cotyledon proteins. On the other hand, in the presence of PMSF, the content of the polypeptides with higher molecular mass including the 97.4 kDa polypeptide and the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (55 kDa) decreased. Besides, when applied together with MeJA, PMSF prevented the appearance of the jasmonate-induced polypeptides (JIPs; 69, 60 and 43 kDa) thus suggesting that JIPs are synthesized from aminoacids released during the breakdown of cotyledon storage proteins. This revised version was published online in July 2006 with corrections to the Cover Date.     

花生种子发育和萌发过程中贮藏蛋白的合成和降解

以花生品种汕油5 2 3种子为材料,分离纯化花生球蛋白的41 kD和38.5 kD两种主要亚基及伴花生球蛋白的6 0.5 KD亚基并制备抗体.We stern blot分析表明,3种亚基在花生胚组织分化期的胚轴和子叶中就开始合成,其中60.5 kD亚基是最先在胚轴和子叶中大量合成和积累的贮藏蛋白,41 kD和38.5 kD亚基在随后的发育中积累量不断增加;种子萌发时这3种亚基的降解进程不一样,胚轴和子叶中41 kD和38.5kD亚基的降解均先于60.5 kD亚基.     

Protein Phosphorylation in Polysomes of Pumpkin Cotyledons after Coumarin Treatment

Abstract: The natural compound, coumarin, caused a change in protein pattern and influenced the phosphorylation status of some ribosome-associated proteins of pumpkin seedlings in vivo and in vitro. Low concentrations of coumarin stimulated ribosome-associated protein phosphorylation only in cotyledons but not in roots and stems. Two phosphoproteins whose phosphorylation state was influenced upon coumarin treatment could be isolated and characterized by their relative molecular weight of about 58 and 65 kDa and pl-values at 5.2 and 5.7, respectively. These phosphoproteins are not major constituents of small or large subunits of ribosomes. We did not find any influence of coumarin on phosphorylation of ribosomal proteins S6, LAO and LAI–3.     

花生种皮抗黄曲霉差异基因表达分析

选择高抗黄曲霉花生种质J11与高感种质金花1012为研究材料,利用基因芯片技术开展了抗病差异基因表达分析研究。试验共使用61078组大豆杂交探针。初步结果表明,与金花1012相比,J11种皮共有417个差异基因表达量上升,650个差异基因表达量下降,其中两份种质的种皮过氧化物酶（POD）差异基因表达量存在明显差异。为初步验证基因芯片分析结果,进行了POD活性测定。结果表明,种子接近成熟前,J11种皮的POD活性迅速上升,明显高于金花1012水平;金花1012则变化不明显,与相关基因差异表达量基本一致。     

绿豆子叶衰老期间核酸代谢的变化

萌发绿豆的子叶自然衰老期间,核酸含量降低,RNA降低的幅度比DNA大。电泳分析结果表明,子叶衰老期间细胞核主带DNA明显降低;而迁移慢的卫星带DNA变化不大。在RNA各组分中,18S rRNA从衰老前期就开始降低;25S rRNA和4～5S小分子RNA到衰老后期才缓慢下降。DNase和RNase活性在子叶整个衰老期间都明显升高,是导致核酸含量下降的主要原因。~3H-核苷掺入试验表明,核酸的合成速率在子叶衰老前期有所上升,到衰老后期又降低。poly(A)~ -mRNA含量在子叶开始衰老时明显上升。     

Analysis of Coat Protein of Peanut stunt virus Subgroup II Isolates from Alfalfa Fields in West Iran

Alfalfa fields in three western provinces of Iran were surveyed for Peanut stunt virus (PSV) during 2011 and 2012. Forty‐seven of 115 samples tested (41%) were infected with PSV. Phylogenetic analysis using coat protein (CP) gene sequences showed that the Iranian isolates belong to the subgroup II of PSV. Pairwise identity analysis revealed four groups representing four phylogenetic subgroups. PSV strains in subgroups III and IV are closely related to each other, as supported by the lowest nucleotide diversity, high pairwise nucleotide identity and high haplotype diversity as evidence of a recent population expansion after a genetic bottleneck. Using the maximum likelihood method, amino acid 86S in the CP gene of the Iranian PSV isolates was found to be under positive selection, although the likelihood ratio test statistics is not significant. This is the first report of the occurrence and phylogenetic relationships of Iranian PSV isolates in west Iran.     

萌发绿豆子叶衰老期间蛋白质代谢的变化

萌发绿豆子叶自然衰老过程中可溶性蛋白质含量一直下降;从衰老开始到衰老前期,总游离氨基酸含量明显上升;但游离氨基酸各组分在子叶衰老期间的变化趋势并不相同。~3H-亮氨酸掺入蛋白质试验和多聚核糖体的相对量及其与总核糖体的比值(P/T)测定都证明在子叶衰老前期有蛋白质的新合成。子叶衰老期间。氨肽酶活性明显降低;而以酪蛋白为底物的蛋白水解酶活性却急剧上升,承担着催化蛋白质降解的主要功能。     

花生种皮蜡质和角质层与黄曲霉侵染和产毒的关系

黄曲霉侵染花生的研究表明,种皮破损的黄曲霉毒素含量显著高于种皮完整的,种皮对黄曲霉侵染和产毒起着重要屏障作用。采用氯仿去除种皮蜡质,用KOH或角质酶去除种皮角质层后,种子黄曲霉感染率和黄曲霉毒素含量显著提高。种皮蜡质和角质层同时去除的与种皮破损的黄曲霉感染率和毒素含量差异不显著,表明种皮的抗性成份主要是蜡质和角质层。种皮蜡质含量测定和种皮表面扫描电镜观察表明,蜡质的含量和角质层的厚度与品种的抗性有关。抗性品种种皮蜡质含量显著高于感病品种。种皮蜡质提取物在体外抑菌效果不显著。说明蜡质的抗性作用主要是物理性阻止黄曲霉菌的穿透。     

不同芸豆品种种子发育过程中贮藏蛋白积累研究

以半蔓生型奶花芸豆(Y09)和直立型奶花芸豆(Y06)为材料,通过SDS-PAGE分析研究了开花后种子形成过程中贮藏蛋白(SP)的积累规律.结果表明:2个芸豆品种开花后种子形成过程中,子叶、胚蛋白积累变化趋势基本一致,但其变化幅度存在显著差异,其中,在开花后20d内,半蔓生型芸豆Y09的籽粒蛋白积累量大于直立型芸豆Y06,但在开花后25~40d,Y06籽粒蛋白积累量显著高于Y09;两芸豆品种的子叶和胚均含有丰富的贮藏蛋白(SP)(14.4~97.4kD),其中子叶的41.0、43.9、39.4kD3种蛋白含量较高,约占子叶总蛋白含量(光密度)50%以上,而胚蛋白亚基分布相对均匀;两品种的子叶、胚中蛋白亚基差异较小,只有个别亚基存在差异.     

Protein Body Formation in Cotyledons of Developing Cowpea (Vigna unguiculata) Seeds

The sequence of protein body formation in cotyledonary cellsof developing cowpea seeds has been followed by electron microscopy.Some protein bodies are formed by sub-division of the main vacuolesof the cotyledon cell but most are formed by expansion of cytoplasmicvesicles which may originate from Golgi bodies. Protein depositionis accompanied by a massive proliferation of rough endoplasmicreticulum and Golgi body production.     

One and Two- dimensional Polyacrylamide Gel Electrophoretic Analysis of Seed Polypeptide Composition of Peanut Cultivars

Seed polypeptides from 46 cultivars of peanut (Arachis hypogaea L. ) were compared by SDS-PAGE and two-dimensional pelyacrylamide gel electrophoresis. Arachin, the major seed storage protein of peanut, showed polymorphism. There were four types of arachin pelypeptide pattems. Type Ⅰ consisted of only four major subunits:41 kD,38.5 kD and two of the 18 kD subunits. Type Ⅱ had six major subunits:41 kD,38.5 kD,37.5 kD and three of the 18 kD subunits. Type Ⅲ consisted of 41 kD, 38.5 kD,36.5 kD and three of the 18 kD subunits. And Type Ⅳ consisted of seven major subunits:41 kD,38.5 kD,37.5 kD,36.5 kD and three of the 18 kD subunits. The compositions of conarachin in different cultivars were similar. Amino acid composition analysis of seed protein in 8 peanut cuhivars showed that Type Ⅰ was rich in methionine and cystine.     

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。