转反义trxs基因对小麦种子α-淀粉酶活性的影响

通过PCR检测，转基因小麦植株均出现474bp的目的带，说明反义trxs基因已整合到小麦基因组上。经测定在种子萌动过程中转基因种子的α—淀粉酶活性低于未转基因种子，证明该基因能够正常表达，从而使转基因品种具有抗穗发芽的能力。     

小麦显性矮秆基因Rht3及其衍生系统抗穗萌特性研究

采用近等基因系法及成熟种子室内萌发试验研究了小麦显性矮秆基因Rht3及其衍生系统的抗穗萌特性,并对普通小麦13个显性、半显性、隐性矮秆基因的抗穗萌性能进行了评价。结果表明,Rht3基因及其株高提升的突变衍生系具有显著而稳定的抗穗萌特性。将Rht3基因回交导入不抗穗萌的普通小麦,受体品种成熟种子的a-淀粉酶活性降低了     

水稻穗萌及其调控的研究进展

水稻穗萌是指水稻种子成熟后期尚未脱离母株之前在穗上发芽的现象.穗萌给农业生产特别是水稻制种造成巨大损失,这一现象与品种遗传特性相关,外界环境也同样起着重要的作用.尽管存在大量有关玉米、小麦等禾谷类作物穗萌机理的报道,但有关水稻穗萌的内在机制仍不十分清楚.本文主要从种子含水量、环境温度、水解酶活性、脱落酸和赤霉素含量、品种遗传特性以及主效微效基因的调控等多个方面概括了水稻穗萌的生理和分子机制,同时也总结了农业生产上从育种、栽培、化学药剂和生物技术途径调控穗萌的多种措施.以期为今后水稻穗萌机理研究以及农业生产上防止穗萌发生提供新的参考.     

转反义trxs基因小麦籽粒中ABA和GAs含量与穗发芽的关系

以转反义trxs基因两个纯合株系及其未转基因对照小麦品种‘豫麦18’为材料,采用荧光定量RT-PCR、整穗发芽试验和酶联免疫吸附（ELISA）的方法,对小麦灌浆过程中反义trxs基因表达、穗发芽特性及内源激素ABA和GAs含量进行了测定,探讨转基因小麦籽粒中内源激素ABA和GAs含量与穗发芽的关系。结果表明,在种子形成过程中,反义trxs基因能够正常表达,两个转基因株系中内源trxh基因表达量都明显低于对照;转基因株系籽粒中ABA和ABA/GAs含量显著高于对照,平均分别比对照升高了18.39%和23.47%,而GAs含量在花后15～30d较对照有所降低,平均比对照降低了9.14%;花后20、25、30d,转基因株系的穗粒发芽率显著或极显著低于对照,平均分别比对照下降了49.65%、28.2%、27.23%;相关分析表明,穗发芽率与ABA和ABA/GAs含量均呈显著或极显著负相关,与GAs含量呈正相关。由此推断,反义trxs基因导入抑制了小麦内源同源基因的表达,改变了小麦体内ABA和GAs的平衡,使ABA合成量增加,是导致转基因小麦穗发芽率降低的原因之一。     

比较和分析NtGNL1的反义寡聚核苷酸抑制在烟草三种培养体系中的效果

为探讨反义寡聚核苷酸抑制(antisense oligodeoxynucleotides inhibition)技术在植物材料上的应用, 以已知功能的NtGNL1 (Nicotiana tabacum GNOM-Like1)为目标基因, 寻找反义寡聚核苷酸抑制最适作用体系, 并进一步分析NtGNL1的具体作用。根据目标基因mRNA序列设计反义寡聚核苷酸序列, 商业合成后并将其添加到烟草胚珠、种子和花粉管离体培养的培养基中, 以抑制NtGNL1的表达。结果表明, 将反义寡聚核苷酸引入离体培养系统, 短时间内抑制了目标基因mRNA的表达, 但对胚胎发育过程的影响和种子萌发的抑制都不明显。在花粉离体萌发系统中, 反义寡聚核苷酸抑制能够高效地引起目标基因mRNA的下调。显微缩时对FM4-64染色的花粉管观察发现, NtGNL1的下调能够引起囊泡分布和运输方向的改变。对花粉管膜流相关的5个基因的半定量分析也显示反义寡聚核苷酸进入花粉管后, 导致3个基因的表达下调, 暗示NtGNL1抑制表达会影响花粉管囊泡运输的多个节点。     

转反义TRX s基因对小麦萌发过程中储藏蛋白降解的影响

以转反义TRX s基因豫麦18和对照（豫麦18）为试材，测定了小麦种子萌发过程中胚乳内thiocalsin、苹果酸脱氢酶、谷丙转氨酶的活性以及游离氨基酸含量的变化。结果表明，反义TRX s基因的导入能够增加对thiocalsin和苹果酸脱氢酶的抑制，降低其活性，使储存蛋白更难于被降解，谷丙转氨酶增高的速度减慢，种子氨基酸代谢减弱。说明蛋白质代谢缓慢是转反义TRX s基因小麦抗穗发芽的一个主要原因。     

穗萌抑制剂的抑制作用及其解除途径的探讨

利用赤霉素，硝酸钾溶液处理受穗萌抑制剂负效影响的杂交稻种子，进行发芽跟踪试验 。结果发现：赤霉素溶液能有效地解除穗萌抑制剂对种子发芽的抑制作用，且龙特甫Ａ种子经８５天，威２０Ａ种子经７０天能自然解除抑制。     

矮生菜豆种子胎萌发生特性的初步研究

研究了若干矮生菜豆品种在不同年份和采种设施下种子的胎萌发生特性.试验结果表明:(1)品种间在种子胎萌发生率方面存在显著差异,有的品种不发生胎萌,有的品种其胎萌种子发生频率在20%以上;(2)对于能够发生种子胎萌的品种而言,采种年份和设施对胎萌种子发生率有明显的影响;(3)种子胎萌一般在开花后35d左右开始发生;(4)胎萌种子的发生部位具有明显的规律,即胎萌种子主要发生在植株的下部和豆荚的近喙端.     

转反义TRX s基因对小麦萌发过程中储藏蛋白降解的影响

以转反义TRX s基因豫麦18和对照（豫麦18）为试材,测定了小麦种子萌发过程中胚乳内thiocalsin、苹果酸脱氢酶、谷丙转氨酶的活性以及游离氨基酸含量的变化。结果表明,反义TRX s基因的导入能够增加对thiocalsin和苹果酸脱氢酶的抑制,降低其活性,使储存蛋白更难于被降解,谷丙转氨酶增高的速度减慢,种子氨基酸代谢减弱。说明蛋白质代谢缓慢是转反义TRX s基因小麦抗穗发芽的一个主要原因。     

转入反义ScP5CS基因烟草植株对PEG和NaCl处理的反应

以转入甘蔗脯氨酸合成酶基因ScP5CS反义片段的转基因烟草与野生型烟草为材料,研究PEG6000与NaCl处理对转基因与野生型烟草植株生长及其T0代种子萌发的影响。结果表明,在17mmol/L NaCl与1%PEG6000胁迫下转入ScP5CS反义基因片段的烟草植株矮小,叶片容易发黄,根系发育迟缓,根长缩短;转基因烟草T0代种子在200mmol/L NaCl胁迫时其发芽率明显受到抑制,并随着盐胁迫处理时间达到第55天时,在50mmol/L NaCl胁迫下其叶片严重黄化,而野生型与转入空载体株系则相反。PEG6000处理对转基因烟草T0代种子萌发影响不大。可见,甘蔗ScP5CS基因在烟草中的反义表达,部分抑制了烟草P5CS基因的活力。     

A simple method to control the seed purity of japonica hybrid rice varieties using PCR-based markers

Cytoplasmic male sterility (CMS) by the cms‐bo cytoplasm and its restoration by the nuclear restorer gene, Rf‐1, are used for seed production of japonica hybrid rice varieties. To produce pure hybrid seeds, a prerequisite is to properly manage the seed purity of parental lines, especially CMS lines. In this study, three dominant polymerase chain reaction (PCR)‐based markers (M1, M2 and M3) were developed to detect mutual contamination in seed batches of CMS lines, maintainer lines, restorer lines and hybrids. M1 detected the mitochondrial sequence that was present in the cytoplasm of common japonica varieties and absent in the cms‐bo cytoplasm. M2 and M3 detected the chromosomal sequence related to the Rf‐1 allele in restorer lines and the rf‐1 allele in common japonica varieties, respectively. By the strategic use of these markers, japonica hybrids and their parental lines could be efficiently distinguished from each other. Furthermore, sensitivity tests for the three markers with a series of crude DNA samples prepared from polished grains demonstrated that these markers could detect one contaminating grain among 500 or 1000 grains. Therefore, the bulk PCR analyses with the markers developed here probably make it possible to control the seed purity of japonica hybrids properly by selecting appropriate seed batches of their parental lines quickly and efficiently.     

关于水稻穗芽的生理学研究

利用重组自交系籼稻中-156/谷梅2号304份株系为材料，在田间条件下营造高温、高湿的小气候，以“穗穗芽率”及“粒穗芽率”为指标，筛选易发生穗芽与不易发生穗芽的极端材料，比较研究重组自交系亲本与各供试株系籽粒在穗芽发生期间的生理差异。结果表明，该重组自交系母本谷梅2号比父本中-156易穗芽，在正常成熟过程中籽粒内源GA₁含量水平高于中-156，ABA含量水平低于中-156；谷梅2号籽粒淀粉酶活性也高于中-156。花后15 d至完熟的籽粒灌浆期如遇高温、高湿条件，谷梅2号及易穗芽的株系籽粒内源GA1随雨日渐增，增幅高于中-156及不易穗芽的株系，同时其淀粉酶活性回升，在正常成熟过程中花后15 d后籽粒中淀粉酶活性与日渐降，但其幅度远高于父本中-156及不易穗芽的株系，这种变化可能是易穗芽的亲本与株系对高温、高湿敏感诱发穗上发芽的主要生理基础。     

Comparison of pedigree selection and single seed descent for oil yield in linseed (Linum usitatissimum L.)

Summary Four populations of linseed derived from five parents were advanced from the F₂ to the F₆ generation by SSD and to the F₅ by pedigree selection. These populations were used to compare the efficiency of the two methods in order to provide superior genotypes with respect to grain yield and oil content, i.e. oil yield. The results showed minimum differences between pedigree and SSD lines for grain yield, where in only one cross the SSD lines were significantly superior to the pedigree lines. Since pedigree selection was carried out for both, seed yield and oil content, a positive response to selection was expected. However, early selection for yield, a character with low heritability, was not successful. On the contrary, significant differences for oil content were detected between the two groups of lines in three of the four crosses studied. In these cases the pedigree lines were superior to the SSD lines. These results demonstrate that strict selection in early generations for oil content, a character with comparatively high heritability, is feasible and successful in linseed. However, selection for seed yield should be postponed to later inbred generations. Consequently, in breeding for maximum oil yield of linseed a two-step selection procedure is recommended.Abbreviations PS pedigree selection - SSD single seed descent - TGW thousand grain weight     

影响西瓜F1代种子数量的遗传因素

用种子数量不同、千粒重不同的亲本进行杂交组配，通过对双亲及F1代种子数量的调查，发现F1代种子数量的多少与双亲种子数量关系不大，而与双亲种子千粒重差别的大小关系密切，双亲种子千粒重差别越大，F1代种子数量越少。从而指出选育少籽F1代的方法。     

Accelerated pedigree selection: An alternative to individual plant selection in the normal pedigree breeding method in the self-pollinated cereals

Summary Accelerated pedigree selection (APS) involves initial selection based on the assessment of lines rather than individual plants. These lines can be derived by accelerated generation procedures. Unlike single seed descent (SSD), line selection can begin in an early generation. This minimises the risk of differential mortality or sterility. Brief outlines of APS for spring and winter cereals are given. The length of the APS breeding cycle is shorter than that of either pedigree selection (PS) or SSD. In addition to higher efficiency of selection for yield, which must be offset against the need for greater resources, it is expected that APS should result in valuable genotypes being retained and better selection for desirable combinations of characters.     

水稻丙酮酸激酶基因OsPK1的过表达转基因株系的获得与鉴定

利用农杆菌介导的转化方法研究OsPK1过表达对水稻的影响。将CaMV 35S启动子驱动的OsPK1的全长CDS序列导入到日本晴基因组中。选择3个过表达转基因系作为代表进行鉴定和分析。结果显示,过表达植株在株高、分蘖数、穗长和千粒重四方面接近野生型水稻,结实率略下降,但每穗饱满种子数明显增加。通过定量RT-PCR分析,发现有4个代谢酶在过表达植株嫩叶中表达存在着上调或下调。用GC-MS方法检测糖含量,显示OsPK1过表达株系中葡萄糖、果糖、半乳糖和蔗糖的含量与野生型差异不大。总之,对OsPK1过表达株系的鉴定,有助于更好了解OsPK1的功能,并且每穗饱满种子数明显增加具潜在的应用价值。     

Toward pollen sterility in German chamomile (Matricaria chamomilla)–Conventional breeding approaches of cytoplasmic/genic male sterility and chemical emasculation

Pollen sterility (PS) in mother lines is crucial for hybrid crossings in plant breeding. 21 di- (2x) and tetraploid (4x) accessions of the hermaphroditic chamomile were screened for spontaneous, thermic or gametocide induced PS. Two crossing trials with the diploid accessions 'Bona' (BON) and 'Hungary 2' (HUN2) should specify maternally inherited cytoplasmic male sterility (CMS). Progeny thereof showed increased PS. BON as mother (BH) entailed means of 30 and 22% PS, versus 8% and 20% after reciprocal crossings. Backcrosses of BH with HUN2 in paternal and maternal direction revealed increasing mean PS (24% and 32%), without verification of CMS. A split plot design of 100 individuals treated with four chemical gametocides analyzed PS, seed set, germination rate and whole plant damage. Tri-iodobenzoic acid caused the highest PS (43%). Progeny of two plants (2x, 4x) with degenerated anthers without pollen, showed 100% PS either in tetraploid F₁ and F₂ (4% and 11%) or in diploid F₂ and F₃ (26% and 23%). High temperature (30/28°C day/night) decreased the male fertility to some extent.     

过氧化物酶及种子蛋白分析鉴定甜椒杂交种纯度

用聚丙烯酰胺均匀胶和等电聚焦电泳技术，分析了甜椒种子及种苗的过氧化物同功酶谱和种子水溶蛋白图谱。在过氧化物酶谱带的两个位点ＰＲＸ－１和ＰＲＸ－２上，自交系亲本与Ｆ１杂交种有明显的差异。水溶蛋白的一个位点ＰＲＯＴ亲本与Ｆ１有明显差异。供试甜椒组合的过氧化物酶和蛋白的图谱，分别具有这些位点。     

外源DNA导入小麦后代变异株系的酯酶同工酶分析

将抗白粉病的大麦ＤＮＡ导入感病的小麦品种后，获得了抗白粉病变异后代，对其中抗性稳定的５个株系子粒酯酶同工酶进行了分析，结果，在变异株系的ＥＳＴ酶谱中，检测到供体大麦所具有的酶带（即转基因酶带）和新的杂种酶带，从而证实了大麦基因（或ＤＮＡ片段）导入到受体中，并被受体细胞基因组整合与表达。