特种稻谷蛋白亚基的研究

以陕西红香米、绿香米、紫糯、茉莉香米、黑帅、黑宝为实验材料,采用SDS-PAGE方法在pH为8.9,电流为15mA条件下,对其种子的谷蛋白进行了电泳分析,寻找其特征及其之间的差异。结果表明绿香米、黑帅都具有9条特征谱带;茉莉香米具有7条特征谱带;紫糯具有6条特征谱带;黑宝具有5条特征谱带;红香米具有4条特征谱带。分子量较高、蛋白谱带较多的品系为绿香米和黑帅(6～5条);分子量较高、蛋白谱带较少的品系为茉莉香米(2条)。     

Rice Allergenic Protein and Molecular-genetic Approach for Hypoallergenic Rice

Allergenic proteins with a molecular mass of about 14 to 16 kDa were isolated from a rice salt-soluble fraction based on the reactivity with IgE antibodies from patients allergic to rice. cDNA clones encoding these allergenic proteins were isolated from a cDNA library of maturing rice seeds, and the deduced amino acid sequences showed considerable similarity to wheat and barley α-amylase/trypsin inhibitors, which have recently been identified as major allergens associated with baker’s asthma. An antisense RNA strategy was applied to repress the allergen gene expression in maturing rice seeds. Immunoblotting and ELISA analyses of the seeds using a monoclonal antibody to a 16-kDa allergen showed that allergen content of seeds from several transgenic rice plants was markedly lower than that of the seeds from parental wild type rice.     

水稻发芽期和幼苗前期耐碱性的鉴定方法研究

通过采用4种不同浓度的Na2CO3模拟碱胁迫,以水稻发芽期发芽势和发芽率的相对碱害率以及幼苗前期的根数、根长和苗高的相对碱害率为鉴定评价指标,研究水稻发芽期和幼苗前期耐碱性鉴定的碱胁迫条件。结果表明,水稻发芽势对碱胁迫的反应较发芽率更为敏感;根长对碱胁迫的反应较苗高和根数更为敏感;幼苗前期对碱胁迫的反应较发芽期更为敏感。在0．15％或0．20％Na2CO3胁迫下水稻发芽势和发芽率的相对碱害率平均值、方差和变异幅度较大。且其分布均匀合理,可作为水稻发芽期耐碱性鉴定的碱胁迫条件;而在0．10％或0．15％Na2CO3胁迫下,幼苗前期根数、根长和苗高的相对碱害率以及综合相对碱害率的平均值、方差和变异幅度较大,且分布相对均匀合理,可作为水稻幼苗前期耐碱性鉴定的碱胁迫条件。在碱胁迫下,水稻发芽势、发芽率、根数、根长和苗高的表型值分别与它们自身的相对碱害率呈极显著的负相关;发芽率的相对碱害率与根数、根长和苗高的相对碱害率呈显著的正相关;发芽期的综合相对碱害率与幼苗前期的综合相对碱害率呈显著的正相关;因此,以水稻发芽期的耐碱性可间接判断水稻幼苗前期的耐碱性。     

The Lack of Correlation between ATP Accumulation in Seeds at the Early Stage of Germination and Seed Quality

Seeds of various species were treated with polyethyleneglycolin the absence or presence of AMP and phosphoenolpyruvate, redriedand examined for ATP accumulation at the early stage of germinationand for the rate of germination under suboptimal temperatures.The pretreatments resulted in seeds with various levels of accumulatedATP but in most comparative experiments no correlation was foundbetween the ATP accumulation and the rate of germination. Similarpretreatments with naturally aged seeds led to the same conclusion.The fact that ATP accumulation is a result of ATP synthesisas well as ATP utilization was demonstrated by a protein inhibitionexperiment which showed a possibility to produce seeds withrelatively high ATP accumulation levels at the early stage ofgermination, but with very low germination ability. Key words: AMP, PEP, Polyethyleneglycol, Pretreatment     

化学药剂对水稻种子萌发和幼苗生长的影响

以早丰 11号水稻种子为材料 ,用KNO3、KH2 PO4 、Na2 HPO4 等不同药液浸种 .结果显示 ,早丰 11号种子在适当浓度的KNO3、KH2 PO4 、Na2 HPO4 浸种后 ,发芽势、发芽率比对照有所提高 ,幼苗生长加快 ,表现在根长、发根数、苗高、鲜重比对照生长好 ,并且对水稻秧苗表现有抵抗低温的作用 .     

茉莉酸甲酯对水稻种子萌发和贮藏物质降解的影响

茉莉酸甲酯 (methyljasmonate ,MeJA)对水稻 (OryzasativaL .)种子萌发及幼苗生长的作用表现出“双重性”效应 ,低浓度促进种子萌发和幼苗生长 ,最适浓度为 5× 10 -7mol/L ,高浓度作用相反。MeJA对水稻萌发胚乳酸性磷酸酶、α_淀粉酶、淀粉和盐溶蛋白降解作用也如此 ,但它抑制谷蛋白降解。实验结果表明 ,MeJA促进和抑制水稻种子萌发都与α_淀粉酶活性有关。讨论了MeJA对种子萌发作用与ABA作用的异同     

Effect of Methyl Jasmonate on the Germination and the Degradation of Storage Reserve in Rice Seed

The effects of methyl jasmonate (MeJA, Nippon Zeon Co., Ltd, Tokyo, Japan) on germination and the degradation of storage reserve in rice （Oryza sativa L.） seeds were studied. There were dual effects of MeJA on seed germination and seedling growth of rice, low concentration of MeJA promoted germination and seedling growth whereas high concentration of MeJA inhibited germination and seedling growth. The optimal concentration of promotion was 5×10-7 mol/L. The acid phosphatase activity, α-amylase activity, degradation of starch and salt-soluble proteins were affected same as germination by MeJA. MeJA inhibited the degradation of glutelins, the higher the concentration, the higher the extent of inhibition above 5×10-7 mol/L.The results showed that either promotion or inhibition of germination by MeJA was very close to the effect on the activity of α-amylase. The mechanisms between abscisic acid and MeJA affection on seed germination and seedling growth were compared.     

Types of Amylases in Rice Grains

The subunit structure of phenoloxidase from the larvae of housefly, Musca domestica vicina Maquart, was investigated by urea treatment, SDS-polyacrylamide gel electrophoresis and electron microscopy. Phenoloxidase was dissociated into a single type of subunit (MW 6.5 × 10⁴) by 5 m urea treatment. A similar subunit (MW 6.1 × 10⁴) was also detected by SDS-polyacrylamide gel electrophoresis. Furthermore, four additional protein bands with molecular weights of 4.8 × 10⁴, 3.5 × 10⁴, 2.4 × 10⁴ and 1.2 × 10⁴ were detected by SDS-polyacrylamide gel electrophoresis by extensive incubation of phenoloxidase with SDS. A cylindrical molecular structure was deduced from the electron microscopic analysis. The outside and inside diameter and the height of phenoloxidase molecule were evaluated to be 100, 50 and 70 Å, respectively.     

抽穗开花期耐高温的爪哇稻资源筛选

通过人工气候室鉴定和分期播种试验,对来自于不同国家和地区的95份爪哇稻进行抽穗开花期耐高温特性评价。以综合结实率为指标,通过聚类分析将95份爪哇稻分为耐热能力不同的3类。综合考虑大田分期播种的结实率和人工气候室鉴定结果,筛选出耐高温品种IRAT109 (IRRI编号38563)、260（IRRI编号14888）和L 4-34（IRRI编号13403）,它们可用于水稻耐高温新品种的选育及耐热性遗传研究。     

水稻胚胎发生与萌发早期分离胚中内源激素的变化

以酶联免疫吸附检测技术分析了水稻(Oryza sativa ssp. japonica)分离胚不同发育时期及萌发早期的内源激素含量的动态变化.GA1含量是所测激素中含量最高的.GA1的变化趋势基本上与ABA相反.花后4 d的胚中GA1和ABA的含量最高;花后8 d到18 d,GA1的含量下降,而ABA含量增加.在早期萌发过程中,种子吸涨后2 d的胚中GA1含量迅速上升,而ABA下降.GA1/ABA的最高比值也出现在吸涨后2 d的胚中.iPAs和ZRs的最高含量也出现在开花后4 d的胚中,但随后含量均下降到相当低的水平,并几乎没有变化.研究结果进一步证实了GA1在早期胚胎发生和萌发过程中起重要的作用;推测iPAs和ZRs可能仅在胚胎发生的早期起作用;GA1与ABA含量之间的相对平衡控制着胚胎发育的过程.用分离胚作为测试材料可以避免胚乳等其他组织成分的干扰,从而比较准确地反映了胚的内源激素变化.此外,本研究是首次用4 d的水稻幼胚作为激素含量测定的起始材料.     

水稻胚胎发生与萌发早期分离胚中内源激素的变化

以酶联免疫吸附检测技术分析了水稻(Oryza sativa ssp．iapoica)分离胚不同发育时期及萌发早期的内源激素含量的动态变化。GA1含量是所测激素中含量最高的。GA1的变化趋势基本上与ABA相反。花后4d的胚中GAl和ABA的含量最高;花后8d到18d,GA1的含量下降,而ABA含量增加。在早期萌发过程中,种子吸涨后2d的胚中GA,含量迅速上升,而ABA下降。GA1／ABA的最高比值也出现在吸涨后2d的胚中。iPAs和ZRs的最高含量也出现在开花后4d的胚中,但随后含量均下降到相当低的水平,并几乎没有变化。研究结果进一步证实了GAl在早期胚胎发生和萌发过程中起重要的作用;推测iPAs和ZRs可能仅在胚胎发生的早期起作用;GA1与ABA含量之间的相对平衡控制着胚胎发育的过程。用分离胚作为测试材料可以避免胚乳等其他组织成分的干扰,从而比较准确地反映了胚的内源激素变化。此外,本研究是首次用4d的水稻幼胚作为激素含量测定的起始材料。     

杂交水稻金优63幼苗期SOD和POD特性研究

对杂交水稻金优63幼苗不同时期的根、茎、叶进行SOD同工酶电泳分析,并测定SOD、POD活性。结果表明,自播种后第7天到第13天,幼苗的SOD同工酶在根、茎、叶中有明显的器官特异性,且SOD活性叶 >茎 >根。相同器官不同时期的SOD同工酶电泳谱带条数及SOD活性都有变化,且SOD活性强弱与SOD同工酶电泳谱带中有无Mn-SOD同工酶带有一定的关系。幼苗的POD活性在根、茎、叶中也有明显的器官特异性,茎中POD活性明显高于根和叶,且POD活性变化与SOD活性变化有一定的关系。     

植物种子萌发期耐盐碱性提高技术研究进展

种子萌发是种胚从生命活动相对静止恢复到生理活跃状态的生长发育过程,最易受到外部环境影响,其中盐碱胁迫是最严重的生态限制因素之一。中国盐碱地面积大、分布广,总面积约1亿hm²,占全国土地面积的10%,盐碱逆境下种子萌发受阻已成为我国盐碱地植物天然更新和农林业生产的首要障碍因子。目前,已知植物种子萌发期耐盐碱性提高技术主要有物理方法、化学调控、生物途径和育种技术4个方面,其中物理方法包括种子引发、低温、超声波、微波、电场等物理因素处理;化学调控包括无机、有机外源物质调控;生物途径包括植物促生细菌和真菌共生;育种技术包括耐盐性鉴定筛选和转基因技术。综述了植物种子萌发期耐盐碱性提高技术研究进展,简述了这些技术的特点及不足,并提出了进一步开展工作的建议,旨在为解决我国盐碱地植物种子萌发受阻难题、实现盐碱地低成本规模化的生物修复和盐碱地区农林业生产提供参考。     

用吸胀初期超微弱化学发光的方法快速检测水稻种子活力

用高灵敏度的单光子计数系统探测了不同贮藏年份的水稻 (OryzasativaL .)品种 80 72 2吸胀初期的超微弱化学发光 (ultraweakchemilumines cence,UCL)。观测到水稻种子吸胀初期 ( 0～ 30min)UCL强度与其老化程度呈负相关 ,水稻种子贮藏时间越长 ,萌发率越低 ,UCL的强度越弱 ,水稻种子UCL强度与萌发率呈极显著正相关。用灵敏的能与单线态氧 ( 1 O2 )反应产生化学发光的发光试剂MCLA ( 2 methyl 6 ( p methoxyphenyl) 3,7 dihy droimidazo[1 ,2 a]pyrazin 3 one) ,检测到吸胀初期水稻种子有单线态氧 ( 1 O2 )的生成 ,产生1 O2 的量与其萌发率呈高度正相关。水稻种子吸胀初期单线态氧的生成是超微弱化学发光的重要诱发因素之一。种子吸胀初期UCL的差异有望成为一种快速、定量、无损伤检测水稻种子活力的新方法。     

水稻资源开花期耐热性的全基因组关联分析

水稻在开花期对高温非常敏感,挖掘耐热种质并解析耐热性的遗传机制,有助于水稻的耐热性遗传改良.本研究选取205份国内外种质资源,在抽穗开花期对遇高温的稻穗进行标记,以高温下标记穗的结实率作为耐热指标,结合高密度SNP标记进行全基因组关联分析并初步预测候选基因.结果 表明:不同水稻种质的耐热性差异明显,高温下的结实率最低为...     

外源CO和NO对水稻种子萌发过程中干旱胁迫损伤的缓解效应

选用水稻品种‘Ⅱ优128’种子为材料,以1.0μmol.L-1高铁血红素(Hematin,H)和200μmol.L-1硝普钠(sodium nitroprusside,SNP)分别作为CO和NO供体,采用PEG-6000模拟干旱胁迫,研究外源CO和NO对干旱胁迫下水稻种子萌发和萌发过程中抗氧化能力的影响。结果表明:高铁血红素和硝普钠处理可以显著提高干旱胁迫下水稻种子的发芽率、芽长和根长;同时显著提高种子淀粉酶活性,显著增加其可溶性糖、可溶性蛋白和脯氨酸含量;还不同程度地诱导增强超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和过氧化物酶(POD)的活性,同时降低质膜相对透性和丙二醛(MDA)含量。研究证实,外源CO和NO可通过调节渗透调节物质含量和保护酶活性来有效缓解干旱胁迫对萌发水稻种子造成的氧化伤害,促进种子萌发生长。     

水稻种子贮藏谷蛋白的微细异质性

利用灵敏的等聚焦与SDS－PAGE合的双向电泳分析方法,从水稻（Oryza sativa L.)种子贮藏谷蛋白中至少可以分离为13条酸性和19条碱性多 肽,依据谷蛋白多肽的表现量推测,水稻谷蛋白主要由约6个主效基因控制,肽图谱与N－端氨基酸序列分析可清晰将谷蛋白酸性多肽分为两组,此两组恰好与谷蛋白GluA和GluB两个cDNA克隆组相吻合。     

Continuous-flow determination of an alpha-amylase inactivator in fermentation samples using a ferricyanide reagent

A continuous-flow system has been developed in which pancreatic alpha-amylase is incubated with soluble starch at 37 degrees C. Reducing sugars being delivered at the "steady-state" hydrolysis of starch are dialyzed into a solution of alkaline ferricyanide. Ferricyanide is then reduced at 95 degrees C. The decrease in the absorbance of ferricyanide solution is recorded as "enzyme baseline." When samples containing alpha-amylase inactivator are introduced into the system, hydrolysis of starch is reduced according to the concentration of enzyme inactivator. The amount of inactivator in the samples is deduced from a standard curve of six standard concentrations using the Technicon Autoanalyzer II system manager in connection with a calculator for automated interpolation. This method allows to determine 40 samples/h with high sensitivity and precision (mean = 1.53 mg/liter; C.V. = 1.4%).