利用Cre/lox重组系统获得烟草TA29-Barnase基因工程雄性不育恢复系

结合F1杂种的生产原理,利用Cre/lox重组系统,建立了一种完全有别于Barnase/Barstar恢复途径的基因工程雄性不育恢复系.将TA29-Barnase雄性不育基因表达盒置于两个同向lox位点之间并与Bar基因融合后获得植物表达载体pBinBarloxTABn,通过农杆菌介导的叶盘转化法将其转入Wisconsin 38烟草获得了雄性不育转基因烟草植株.雄性不育转基因植株表现花药瘦小,不饱满,花粉量明显偏少,花粉畸形、皱缩、塌陷、极少萌发,自交无法正常座果结子.TA29-Barnase雄性不育转基因植株用来自pBinCre转基因植株的花粉授粉后则能正常结果结子.分别从pBinCre转基因植株C1与转基因雄性不育植株BN1、BN6杂交后代中筛选到23株、22株Kan和PPT双抗性植株,分子检测结果显示两个杂交组合后代中凡同时含Cre基因和Bar基因的F1植株,其TA29-Barnase雄性不育基因均被删除,删除效率达到100%.F1植株中的TA29-Barnase基因被删除后,育性被恢复,能正常开花结果.     

重离子辐射诱导玉米雄性不育突变系的遗传研究

利用30Gy7Li离子辐射玉米杂交种农大108的种子,从M3代材料中获得雄性不育突变材料39I3-2,并通过套袋杂交获得了不育材料的杂交组合。调查不育突变植株性状及其杂交组合和F2代雄花育性,结果表明,不育材料39I3-2花粉败育彻底,不育性状表现稳定,呈现出由隐性单基因控制的核不育的遗传特点。雄性不育突变体的出现与重离子辐射诱导有关,为基因突变的结果。     

辐照花粉辅助授粉对百合远缘杂交结实的影响及杂种早期分子鉴定

研究了辐照花粉辅助授粉对百合‘pollyanna’(♀)×毛百合杂交结实的影响。结果表明:采用普通柱头授粉能获得许多膨大果实和饱满种子,表明该组合为亲合性组合。用经1000和2500Gy的60Coγ射线辐照的母本花粉进行辅助授粉明显提高了果实中饱满种子的结籽率,表明2种剂量的辐照花粉辅助授粉均能有效克服该组合的杂交障碍。结籽数的提高能获得更多的后代,增加了从F1代中选育新优单株的几率。随机选取辐照花粉辅助授粉获得的杂交子代15株,采用ISSR技术对其组培苗进行亲子鉴定,6条ISSR随机引物在15个子代中共扩增出372个条带,其中具有父本特征带114条,具有母本特征带67条,具有双亲共有带65条。ISSR鉴定结果证实了杂种的真实性。     

籼稻体细胞无性系雄性不育突变体后代遗传及其恢保关系鉴定

对野败型保持系珍汕97B、恢复系IR24、IR26和泰引1号等4个水稻材料的幼穗在不同的培养基上培养、再生植株及对其后代进行育性和测交鉴定及其它遗传分析,探讨了雄性不育变异株的恢保关系,结果表明:在IR26、泰引1号和珍汕97B的R1、R2群体中都获得了不同比率的雄性不育变异株,这些不育株的花粉败育可分为无花粉、典败、圆败和染败4种类型.对离体培养产生的雄性不育变异株用一批现有CMS不育系的保持系和恢复系进行杂交,其中有2株分别来自IR26(编号为269157)和珍汕97B(编号为B29154)的不育株,其恢保关系与野败型一致,现已回交5代.来自IR26(编号为2692411)的另一雄性不育株,用上述亲本已连续回交5代,后代均不育,其恢保关系似与野败不同.而有些来自珍汕97B和IR26的12株雄性不育变异株,杂交后代(F1)全部可育,结实率85%以上.     

2个马铃薯杂交组合F1的SSR鉴定

杂交育种是马铃薯新品种选育的重要方法, 其杂种F₁真实性鉴定是获得目标性状单株的关键环节。为选育优质、高产、抗病性及抗旱性强的马铃薯新品种, 用马铃薯品种"费乌瑞它"(Favorita)分别与"J07-6"和"陇薯3号"杂交, 获得了杂种F1代。本试验利用SSR标记技术对"Favorita"×"J07-6"、"Favorita"×"陇薯3号"2个杂交组合F1共86个单株的真实性进行了鉴定。试验从43对SSR引物中筛选出2对适宜引物STM1049和S7。利用这2对引物进行PCR扩增, 将"Favorita"×"J07-6"杂交种F1和"Favorita"×"陇薯3号"杂交种F1的SSR带型划分为双亲互补型、缺失型、父本型和母本型4种类型。依据SSR带型特征, 从"Favorita"×"J07-6"和"Favorita"×"陇薯3号"2个杂交组合F₁单株中分别鉴定出真杂种34个和27个, SSR分子标记技术用于马铃薯杂交种真实性鉴定是可靠的。该研究结果可为马铃薯杂交种优良株系选育提供依据。     

油茶无性系组织培养繁育实验

组织培养作为无性系培育繁殖的重要方式,具有缩短育种时间,节省母本资源、子体抗性强等明显优势,因此对油茶进行无性系组培对于提高油茶育种质量以及种质资源的普及发挥着关键性的作用。基于此,从油茶芽和愈伤组织诱导试验的角度探讨了油茶无性系组织培养的相关问题,为保存油茶种质资源的优良性状并实现大范围的推广提供技术支持。     

辐射水稻雄性不育性在异源胞质背景的遗传表达

60 Coγ射线辐射水稻恢复系获得 4个隐性单基因不等位雄性不育突变体。用雄性不育突变株 可育株杂合体植株作父本 ,与 1 5个种质的同核异质代换系 (作母本 ) ,杂交配组成 60个组合。结果F1代小穗正常可育 ,F2 代分离出数量不等的雄性不育株。未发现雄性不育基因与异源胞质互作表现完全可育的基因型。     

化学杂交剂诱导的雄性不育花药组织内源激素的变化

实验以Ｋ７８－１Ｂ和Ｔ０００２Ｂ为材料，分别用化学杂交剂ＳＣ２０５３和ＢＡＵ２诱导完全和部分雄性不育，测定了花粉败育过程中花药组织各种内源激素的变化。结果表明花药组织内源ＩＡＡ、ＤＨＺＲ水平都低于对照，ＺＲ和ＡＢＡ含量高于对照，变化幅度与喷施杂交剂的量密切相关，ＩＰＡ与ＧＡｓ的变化因小麦基因型，杂交剂种类及剂量而异，这说明化学杂交剂可能了内源激素的含量从而导致了雄性不育的发生，有助于进一步阐明化学     

辐射诱发瓜叶菊雄性不育系及其利用研究

本研究用６０Ｃｏγ射线５０Ｇｙ辐照１、２年生瓜叶菊的于种子，获得了光枝叶、部分花粉败育的突变体，然后自交分离出花粉败育率１００％的雄性不育系。其突变率为２．４％，突变雄性不育是由隐性基因控制、基因突变产生的。１９８７年以来，先后诱发获得瓜叶菊雄性不育系蓝花１、２、３和玫瑰红花４、５号。其中１、２、３号与瓜叶菊自支系杂交，育成了Ｆ１代杂种，并具有一定的杂种优势。     

1份~(60)Co-γ射线诱变玉米雄性不育突变体的遗传分析

为了研究一个新玉米雄性不育突变体的遗传模式,将~(60)Co-γ射线诱变选育的玉米K305雄性不育突变体姊妹交群体在不同地点、不同年份、不同季节种植,采用室内花粉镜检与田间调查相结合的方法进行育性鉴定。通过不育株姊妹交、测交及测交组合的自交和回交后代,进行细胞核效应分析;以具有正常细胞质的自交系为母本,育性完全恢复的测交种及姊妹交群体中的可育株为父本进行反交,对其反交的F1及F2进行细胞质效应分析。结果表明,不育株雄穗无花药外露,花药干瘪,败育彻底,不育性状稳定;姊妹交和回交后代育性分离比例为1∶1,自交后代育性分离比例为3∶1,不育性状表达与细胞质的改变无关。说明该突变体是由隐性单基因控制的可遗传的"无花粉型"细胞核雄性不育,为其进一步开发利用奠定了基础。     

旱稻(Oryza sativa)×长芒稗(Echinochloa caudata)远缘杂交后代结实率及杂种优势分析

以旱稻基因型旱 65(OryzasativaL .)为母本 ,稗草长芒稗 (Echinochloacaudata)为父本进行远缘杂交 ,获得实粒种子。连续 5年试验结果表明 :( 1 )F0 高度不孕 ,并伴有杂交种发育夭折现象。去雄 82 4朵小花 ,结实率为 1 2 1‰。 ( 2 )F1杂种优势明显 ,育性正常 ,结实率 92 2 6% ,单株产量超母本 80 3 9%。 ( 3 )F2 农艺性状分离严重 ,但育性无分离。群体 40 5株无不育株出现。选育出的优良变异单株“远F2 1” ,超亲优势为 55 64%。 ( 4)对“远F2 1”衍生F3株系 1 60株考种株穗数、株高、穗长、主茎穗一级枝梗数、穗总粒数、穗实粒数诸性状 ,平均超亲优势分别为 2 3 2 %、1 5 1 8%、47 2 6%、3 3 84%、2 4 2 6%及 2 1 62 % ,差异显著。 ( 5)F5光合速率测定 ,超母本2 9 72 %。远缘杂交对不同属间引入目的基因 ,非常有效。     

Inventory methods for finding historically cultivated hop (<Emphasis Type="Italic">Humulus lupulus</Emphasis> L.) in Sweden

The relationship between the conservation of plant genetic resources and their culture history is intimate. Consequently, biodiversity research must also take into account historical and cultural factors. An inventory of plants collected from all over Sweden was made with the aim of establishing a national Swedish gene bank for once cultivated hop. Only female hop, which could be regarded as cultivated hop on the basis of history, were selected. In this study, two different inventory methods were used: one based on plant habitat and the other involving the use of historical documents, primarily large-scale maps from the first half of the seventeenth century, used to locate their cultivars today. The documented history of the hop, combined with its biology, is the basis for the methods used. Hop is the only Swedish crop that according to a 1442 law had to be cultivated. The law lasted for over 400 years. Since the hop is a perennial, dioecious plant and only female individuals are cultivated, over time very few genetic recombination events are expected. Today, it is possible to connect and identify living plants using historical documents. The degree of connection between today’s living plants and the historical evidence for hop cultivation differ between the two methods.     

Genetic Engineering in Pea Crop Improvement

Abstract

Genetic engineering can be used to complement traditional breeding programmes in those plant species where suitable tissue culture and transformation methods allowing regeneration of fertile transgenic plants have been developed. Pea, an important legume crop, was earlier considered to be recalcitrant to genetic engineering. Recent advances in tissue culture methods have, however, made it possible to obtain fertile transgenic pea plants. This paper summarizes the results obtained in developing transformation methods for pea.     

In vitro propagation and germplasm cold-storage of fertile and male-sterile Allium trifoliatum subsp. hirsutum

Summary Procedures for in vitro clonal propagation and for cold-storage of propagules were developed for fertile and male-sterile genotypes of Allium trifoliatum subsp. hirsutum, var. hirsutum and var. sterile, respectively. Highest rate of shoot multiplication was achieved from basal leaf and umbel explants on a modified BDS medium supplemented with 9 mg/l benzyladenine. Naphthalene acetic acid reduced the propagation rate, and was not required for shoot multiplication. The resulting shoots were rooted in an indole butyric acid-supplemented medium, and bulbing occurred upon exposure to a 16 h photoperiod. The small dormant bulbs were transplanted into potting mixture and sprouted after termination of dormancy, resulting in phenotypically-normal plants. No significant differences, in either shoot regeneration or plant establishment, were found between fertile and male-sterile genotypes.Basal leaf explants of in vitro grown plantlets were stored at 4–6°C for up to 16 months. Standard medium, or modified media containing 0.4 to 10.0% sucrose or 1 to 10 mg/l paclobutrazol, were used in storage experiments. Eighty to 100% and 70% survival rates, were recorded after 8 and 16 months cold-storage, respectively, in a 10% sucrose medium. Following their transfer to standard culture conditions, these explants regenerated plants which were phenotypically similar to those developed in the potting mixture.Isozyme polymorphism of 13 proteins was used to assess the genetic stability of cold-stored explants. Considerable differences in zymograms were found between the fertile and the sterile varieties. However, no differences in isozyme profiles were detected between the control field-grown plants, and those which were established from in vitro-stored leaf base explants. The only exception were plantlets exposed to a high paclobutrazol concentration in storage. The latter exhibited a doubling in the alcohol dehydrogenase profile.Abbreviations ADH Alcohol dehydrogenase - BA benzyl adenine - IBPGR International Board for Plant Genetic Resources - MDH malate dehydrogenase - MS Murashige & Skoog medium - NAA naphthalene acetic acid     

Detection of nifH Sequences in Sugarcane (Saccharum officinarum L.) and Pineapple (Ananas comosus [L.] Merr.)

To detect N₂-fixing bacteria in a plant without using culture methods, nifH gene segments were amplified with degenerate primers from DNA extracted from stems of sugarcane ( Saccharum officinarum L.) and leaves of pineapple ( Ananas comosus [L.] Merr.). Sequences of the nifH clones were homologous to those of bacteria in the genera Bradyrhizobium, Serratia , and Klebsiella. The bacteria with nifH sequences homologous to those of the three genera may be promising candidates for predominant endophytic diazotrophs in sugarcane and pineapple, because these sequences were commonly detected in samples from different host plants, which were collected at different locations. On the other hand, no nifH sequence related to Gluconacetobacter diazotrophicus , which is an endophytic N₂-fixing bacterium and had been previously isolated from sugarcane, was detected in sugarcane. This indicates the absence or the presence of few G. diazotrophicus in the stems of the sugarcane plants used in the current study.     

不同生长环境下黄顶菊浸提液对多年生黑麦草萌发与生长的影响

为阐明黄顶菊入侵机制,本文在调查不同生境下黄顶菊居群特征的基础上,进一步采用植株浸提液生物测试的方法研究了不同生境、生长密度及替代植被下黄顶菊对多年生黑麦草的化感潜力。结果表明:黄顶菊的化感作用强弱与其入侵能力存在一定相关性,黄顶菊浸提液的化感潜力随浓度升高而增强;不同植株组织浸提液的化感潜力存在显著差异,表现为地下部分明显低于地上部分;不同生境下黄顶菊浸提液的化感潜力顺序为水边>果园>路边,50 g.L 1茎叶浸提液对多年生黑麦草发芽率化感效应敏感指数分别为2.83(水边)、1.31(果园)和0.71(路边);不同密度黄顶菊的茎叶化感潜力顺序为高、中密度区显著大于低密度区,50g.L 1浓度浸提液对多年生黑麦草发芽率化感效应敏感指数分别为3.91(高密度)、4.89(中密度)和0.29(低密度);根部浸提液的化感潜力顺序则相反。不同替代植被下茎叶浸提液的化感效应为混种>单种,而根系浸提液的化感效应敏感指数则差别不大。因此,黄顶菊不同组织器官化感活性物质分配不同,同时其化感效力能够适应环境条件而发生变化,这可能是造成黄顶菊在不同环境中入侵效果差异的原因之一。     

Iron‐manganese interactions among clones of Nilegrass

Tetraploid clones of Nilegrass (Acroceras macrum, Stapf.) develop a chlorosis resembling iron (Fe) deficiency on acid (pH 5.0) soils in the Midlands of KwaZulu, Natal, South Africa. Hexaploid and pentaploid clones appear more resistant to the disorder. Iron deficiency would not be expected in such acid soils, but foliar sprays of Fe sulfate reduce the symptoms within 24 hours. Aluminum (Al) toxiciry has been ruled out as a cause of this chlorosis on the basis of soil tests. Manganese (Mn)‐induced Fe deficiency has been postulated. Six Nilegrass clones, differing in ploidy levels, were grown under low Fe or high Mn levels in nutrient solutions, in Mn‐toxic soil, in calcareous soil and in a standard potting soil at pH 7.0. Differential chlorosis symptoms, similar to those observed in the field, were reproduced in plants grown in low Fe or high Mn solutions, in neutral potting soil and in calcareous soil at pH 7.8. Based on plant symptoms and dry weights, the tetraploids were generally more sensitive to these conditions than hexaploid or pentaploid clones. However, in Mn‐toxic soil, plants had leaf tip necrosis rather than the chlorosis typical of Fe deficiency. When grown in a standard potting soil at pH 7.0, plants showing chlorosis accumulated higher concentrations of phosphorus (P), Al, copper (Cu), Mn, Fe, and zinc (Zn) than non‐chlorotic plants. Differential susceptibility to chlorosis is apparently associated with interference of such elements in Fe metabolism, and not with differential Fe concentrations in plant shoots. Additional studies are needed to determine the chemical states of Fe and Mn in root zones and within plant shoots of these clones. Resolution of the differential chlorosis phenomenon would contribute to fundamental knowledge in mineral nutrition and could be helpful in tailoring plant genotypes to fit problem soils.     

Flumequine Uptake and the Aquatic Duckweed, Lemna minor L.

Phytotoxicity of Flumequine (F) on the aquatic duckweed, Lemna minor L., and plant drug uptake were evaluated by a simple ecotoxicological test. Flumequine, at all concentrations between 50 and 1000 μg L^-1 tested, affected the plant growth: leaves and roots were damaged, but duckweed continued to grow on a five weeks period. Furthermore, increasing drug concentrations decreased the chlorophyll b content in plants. These effects depend on F uptake by plants, which is quite high (from 0.72 to 13.93 μg g^-1 plant dry weight). Based on this activity, Lemna can be taken into consideration as a tool for in situ remediation of drug contaminated waters: the presence of Lemna significantly lower the F concentration in culture media on a five weeks period. Results strongly support its remediation capability.     

Manganese content of the rice plant under water culture conditions

The rice plant has been known to have a high tolerance to manganese. Tissues of rice plants have been found to contain as much as 4,000 ppm Mn with no apparent harmful effects (1). On the other hand, others have reported manganese toxicity in tissues containing 2,000 ppm Mn (2, 3). Under water culture conditions, 10 ppm Mn was reported to be the upper critical level in the culture solution for maintaining normai rice growth; at this level, the straw contained 1,000 ppm Mn (4). Toxicity symptoms started to develop at 30 ppm Mn in culture solution (5).     

Uptake, Deposition and Wash Off of Fluoride and Aluminium in Plant Foliage in the Vicinity of an Aluminium Smelter in Norway

Uptake, deposition and wash off of fluoride and aluminium in/on plant foliage were studied in two experiments near an Al smelter by growing different plant species under cover with free wind-flow underneath, and in the open, and by washing the leaves. With an average ambient air concentration of 2.2 μg F m³, the F and Al contents in leaves varied between species from 138 to 665 mg F kg^?1 and from 150 to 1025 mg Al kg^?1when grown under cover. An average of about 60% of the F and nearly 70% of the Al could be washed off covered plants. Uncovered plants had 33–51% lower F content, indicating a considerable precipitation wash off. Precipitation wash off varied with species, probably due to different leaf surface properties. The strong effect of precipitation on plant foliage F has to be taken into consideration when comparing differences in F content between years, localities and species. On average about 40–50% of the F and 55% of the Al could be washed off the leaves of uncovered plants. Washed leaves of plants exposed to precipitation had about 20% lower F content, compared to washed leaves of covered plants, thus indicating leaching of F from leaves exposed to rain. In contrast to fluoride, the Al contents in washed leaves of uncovered plants were about 50–80% higher than in covered washed leaves, thus indicating a significant uptake of Al solved in rainwater through the leaf cuticle.