大豆(Glycine max(L.)Merr.)致敏蛋白Gly m Bd 30K检测方法及低致敏优异种质鉴定

Gly m Bd 30K蛋白是大豆中主要的免疫显性过敏原之一,会引起人和牲畜腹泻和肠道炎症等过敏反应.因此,发掘低Gly m Bd 30K蛋白含量优异种质对于培育优质大豆品种具有重要意义.为了获得致敏蛋白Gly m Bd 30K低含量的优异种质,根据Gly m Bd 30K蛋白的190-379aa多肽序列制备多克隆抗体...     

Sampling DNA from the rhizosphere of Brassica napus to investigate rhizobacterial community structure

Variations in genotype rankings among screenings for Al tolerance in hydroponics may be related to differences in the composition of the solutions. In the present study, we investigated the involvement of Mg ions in modifying Al rhizotoxicity in soybeans. Root elongation was strongly inhibited by Al in a simple, 800 M CaSO₄ solution, but elongation increased noticeably when the solutions also contained Mg. Amelioration of Al rhizotoxicity was not associated with an increase in ionic strength of treatment solutions because Al³⁺ activities were kept constant. Concentration series experiments indicated that the Mg effect occurred in the M range, while Ca amelioration of Al toxicity occurred at mM concentrations. The positive effect of Mg on root elongation was greatest for Al-sensitive genotypes and minimized genotypic differences for Al-tolerance. The Mg protection against Al rhizotoxicity apparently does not occur with all species, because it was not observed in Atlas and Scout 66 wheat varieties. The ability of Mg to ameliorate Al toxicity in soybean at M levels suggests the involvement of distinct physiological factors.     

The composition of newly synthesized proteins in the endoplasmic reticulum determines the transport pathways of soybean seed storage proteins

Glycinin (11S) and beta-conglycinin (7S) are major storage proteins in soybean (Glycine max L.) seeds and accumulate in the protein storage vacuole (PSV). These proteins are synthesized in the endoplasmic reticulum (ER) and transported to the PSV by vesicles. Electron microscopic analysis of developing soybean cotyledons of the wild type and mutants with storage protein composition different from that of the wild type showed that there are two transport pathways: one is via the Golgi and the other bypasses it. Golgi-derived vesicles were observed in all lines used in this study and formed smooth dense bodies with a diameter of 0.5 to several micrometers. ER-derived protein bodies (PBs) with a diameter of 0.3-0.5 microm were observed at high frequency in the mutants containing higher amount of 11S group I subunit than the wild type, whereas they were hardly observed in the mutants lacking 11S group I subunit. These indicate that pro11S group I may affect the formation of PBs. Thus, the composition of newly synthesized proteins in the ER is important in the selection of the transport pathways.     

Efficient<Emphasis Type="Italic"> Agrobacterium tumefaciens</Emphasis>-mediated transformation of soybeans using an embryonic tip regeneration system

Here, we report the establishment of an efficient, in vitro, shoot organogenesis, regeneration system for soybeans [Glycine max (L.) Merr.]. Mature soybean seeds were soaked for 24 h, the embryonic tips were collected and cultured on MSB5 medium supplemented with 3.5 mg l^–1 N⁶-benzylaminopurine (BAP) for 24 h, and explants were transferred to MSB5 medium supplemented with 0.2 mg l^–1 BAP and 0.2 mg l^–1 indolebutyric acid. Use of embryonic tips yielded a higher regeneration frequency (87.7%) than regeneration systems using cotyledonary nodes (40.3%) and hypocotyl segments (56.4%) as starting materials. Regenerated embryonic tips were inoculated with Agrobacterium tumefaciens strain EHA105, which contains the binary vector pCAMBIA2301, and cultured for 20 h. Our results showed that the T-DNA transfer efficiency reached up to 78.2% and the transformation efficiency reached up to 15.8%. These data indicate that the embryonic tip regeneration system can be used for efficient, effective Agrobacterium-mediated transformation.Abbreviations GUS -Glucuronidase - T-DNA Transferred DNA - BAP N⁶-Benzylaminopurine - IBA Indolebutyric acid     

Ectopic expression of a soybean phytase in developing seeds of <Emphasis Type="Italic">Glycine max</Emphasis> to improve phosphorus availability

A transgenic approach was used to alter soybean seed phytate content by expressing a soybean phytase gene (GmPhy) during seed development to degrade accumulating phytic acid (IP₆). An expression vector containing the soybean phytase cDNA controlled by the seed-specific -conglycinin promoter (-subunit) was used to transform embryogenic soybean cultures. Plants from four independent transgenic lines were analyzed for transgene integration and seed IP₆ levels. The reduction in IP₆ levels in transgenic seeds compared to control Jack soybeans ranged from 12.6 to 24.8 as determined by HPLC. A low copy transformant was propagated to the T₄ generation and examined in more detail for phytase expression and enzyme activity during seed development. Expression of phytase mRNA and phytase activity increased during seed development, consistent with the use of an embryo-specific promoter. Ectopic phytase expression during seed development offers potential as an effective strategy for reducing phytate content in soybean seed.     

Isolation and characterization of a root nodule-specific cysteine proteinase cDNA from soybean

We have determined that a nodule-specific cDNA clone (GmCysP1), obtained from a soybean root nodule-specific EST pool, encodes cysteine proteinase. Its amino acid sequence homology, as well as the conservation of typical motifs and amino acid residues involved in active site formation, shows that GmCysP1 can be classified as a legumain (C13) family cysteine proteinase, belonging to clan CD. Moreover, based on its expression patterns,GmCysP1 is a nodule-specific cysteine proteinase gene that is possibly associated with nodule development or senescence. Our genomic Southern analysis also suggests thatGmCysP1 is a member of a multigene family. Therefore, we propose that GmCysP1 is the first to be identified as a nodule-specific and senescence-related cysteine proteinase that belongs to the legumain family from soybean.     

Production and purification of recombinant DP1B silk-like protein in plants

Spider dragline silk of Nephila clavipes consists of two highly repetitive proteins, spidroin 1 and spidroin 2. To develop a plant platform for production of recombinant silk-like protein, two plant-optimized DP1B genes were synthesized to mimic spidroin 1. DP1B-8P encodes for a 64 kD DP1B silk-like protein and DP1B-16P for a 127 kD DP1B silk-like protein. Both genes have been introduced into Arabidopsis for leaf-based production driven by the 35S promoter and for seed-specific production driven by the -conglycinin subunit promoter, respectively. They have also been expressed in somatic soybean embryos under the control of the -conglycinin subunit promoter. The results demonstrate the synthesis and accumulation of DP1B silk-like protein in leaves and seeds of Arabidopsis, as well as in somatic soybean embryos. They suggest that seeds are the more preferred tissue for DP1B production since they offer higher production yield and quality. In addition, a simplified protocol for purifying DP1B from plant tissue is discussed.     

低能离子注入对大豆种子吸胀冷害的影响

本文分别从浸种液中无机离子的浓度、可溶性糖的浓度以及溶液的pH值,研究了低能离子注入对大豆种子吸胀冷害的影响。结果发现一定剂量的氮离子注入大豆种子后,其无机离子、可溶性糖、酸性物质的泄漏量均有低于对照组的趋势,而且其长势好于对照组,说明离子注入能在一定程度上减轻大豆遭受的吸胀冷害。     

3,4-Dichloroaniline is detoxified and exported via different pathways in Arabidopsis and soybean

The metabolic fate of [UL-14C]-3,4-dichloroaniline (DCA) was investigated in Arabidopsis root cultures and soybean plants over a 48 h period following treatment via the root media. DCA was rapidly taken up by both species and metabolised, predominantly to N-malonyl-DCA in soybean and N-glucosyl-DCA in Arabidopsis. Synthesis occurred in the roots and the respective conjugates were largely exported into the culture medium, a smaller proportion being retained within the plant tissue. Once conjugated, the DCA metabolites in the medium were not then readily taken up by roots of either species. The difference in the routes of DCA detoxification in the two plants could be explained partly by the relative activities of the respective conjugating enzymes, soybean containing high DCA-N-malonyltransferase activity, while in Arabidopsis DCA-N-glucosyltransferase activity predominated. A pre-treatment of plants with DCA increased DCA-N-malonyltransferase activity in soybean but not in Arabidopsis, indicating differential regulation of this enzyme in the two plant species. This study demonstrates that DCA can undergo two distinct detoxification mechanisms which both lead to the export of conjugated metabolites from roots into the surrounding medium in contrast to the vacuolar deposition more commonly associated with the metabolism of xenobiotics in plants.     

Polymerization of cardanol using soybean peroxidase and its potential application as anti-biofilm coating material

Soybean peroxidase (20 mg) catalyzed the oxidative polymerization of cardanol in 2-propanol/phospate buffer solution (25 ml, 1:1 v/v) and yielded 62% polycardanol over 6 h. Cobalt naphthenate (0.5% w/w) catalyzed the crosslinking of polycardanol and the final hardness of crosslinked polycardanol film exceeded 9 H scale as pencil scratch hardness, which shows a high potential as a commercial coating material. In addition, it showed an excellent anti-biofouling activity to Pseudomonas fluorescens compared to other polymeric materials such as polypropylene.