转抗虫基因三倍体毛白杨植株体内农杆菌残存与逃逸

用部分改造的BtCry1Ac基因与慈菇蛋白酶抑制剂（APLA）基因构建的双抗虫基因表达载体,通过农杆菌介导法对三倍体毛白杨进行了转化,对转化后植株体内残存农杆菌在继代培养和移栽过程中进行了跟踪检测。结果表明：通过对转化再生植株的分子生物学检测,42个株系中,33个株系为阳性,阳性率达到80％;用Bt毒蛋白抗血清进行ELSA检测结果表明,7个转基因株系都有Bt杀虫蛋白表达;基因转化后,可采用附加50mg／L卡那霉素,300mg／L羧苄青霉素的筛选培养基消除细菌并进行抗性芽筛选。对28个转基因株系叶片、茎段和根段在含有卡那霉素50mg／LYEB培养基上进行细菌培养,通过在T-DNA区、质粒Vir区和农杆菌基因组设计引物,进行PCR检测,证明有3个株系（33、37、5号）检测到残存工程农杆菌,并在组培瓶中存活24个月。将带菌的3个株系组培苗移栽到花盆中,室内培养1个月后,在33号株系根际土壤中检测到了目的农杆菌。     

芥菜型油菜抗虫转基因植株及其后代株系的研究*

带有1～2mm子叶柄的芥菜型油菜子叶经农杆菌感染后,培养在附加10～20mg/L卡那霉素的MS选择培养基上筛选转化愈伤组织及不定芽。卡那霉素抗性苗相继在含30～50mg/L卡那霉素的选择培养基上继代培养,再转移到含20mg/L卡那霉素的生根培养基上诱导生根。以苏云金杆菌杀虫晶体蛋白基因为探针,进行Southern blot分子杂交,得到阳性结果。PCR分析也证明外源基因整合到油菜基因组并稳定传递到后代。转基因植株的抗虫性和卡那霉素抗性在自交后代中得到保持,筛选得到纯合的转基因植株后代株系。     

转双抗虫基因三倍体毛白杨外源基因表达及性状相关性分析

用部分改造的BtCry1Ac基因与慈菇蛋白酶抑制剂（API）基因构建的双抗虫基因表达载体,通过农杆菌介导法转化了三倍体毛白杨Populus tomentosa Carr.,获得一批转双抗虫基因株系。对转基因株系的抗虫毒蛋白表达进行了ELISA和Western Blot检测,同时用转基因株系叶片对杨扇舟蛾Clostera anachoreta Fabricius和舞毒蛾Lymantria dispar L.幼虫进行室内饲虫试验,并对各项外源基因表达指标进行了相关分析。结果表明：在检测的28个转基因株系中,对杨扇舟蛾高抗株系占总参试系号的41％,中抗系号占35.0％,低抗系号占24％,对舞毒蛾高抗株系占参试系号的70％。转基因植株可明显抑制存活幼虫的生长发育,且不同转基因株系饲养的幼虫发育存在显著差异。连续两年相关分析表明,不同转基因株系幼虫死亡率间存在极显著相关。转基因植株对舞毒蛾和杨扇舟蛾均表达出抗虫性,并存在极显著相关。ELISA检测结果表明,不同转基因株系Bt毒蛋白表达量存在差异,变化在0.0011％～0.0161％。转基因植株对害虫的杀虫效果与Bt杀虫蛋白的表达量存在显著相关,表明Bt毒蛋白在抗虫效果中占有重要地位。转基因植株对卡那霉素表现出一定的抗性,但与抗虫程度相关不明显,单纯用卡那霉素作为筛选手段并不能完全反映植株的真实抗虫效果。     

ACC合成酶基因及其反义基因对西瓜的遗传转化

以2日龄西瓜(Citrullus lanatus(Thunb.)Mansfeld)无菌苗子叶为外植体,通过与根癌农杆菌(Agrobacterium tumefaciens)进行叶盘共培养建立了西瓜的遗传转化系统。所用根癌农杆菌中含有改建后分别携带嵌合NPTⅡ基因和番茄的ACC合成酶基因及其反义基因的质粒。外植体在MSA培养基(MS盐类、B_5维生素、1.0mg/L BA、0.2mg/L IAA)上预培养3～4d后,与根癌农杆菌共培养4d,随后转移外植体至附加100mg/L卡那霉素、300mg/L头孢菌素的MSA培养基上筛选转化芽。将带芽外植体移入含有100mg/L卡那霉素、300mg/L头孢菌素的伸长培养基(MS 0.2mg/L KT)上进行芽伸长,切取2～3cm高的伸长芽移入生根培养基(1/2MS 0.1mg/L NAA)生根。Southern blot结果证明获得转基因植株,乙烯释放指标表明转入的正义和反义ACC合成酶基因得到不同程度的表达。     

转cry1Ab基因抗虫水稻的田间试验

采用农杆菌介导的遗传转化技术, 将Bt基因cry1Ab导入三系恢复系明恢63获得了一批抗虫性及农艺性状较好的转基因纯合株系。研究中, 进一步检测了这些纯合株系的拷贝数、Bt蛋白含量、抗虫性及主要农艺性状。结果显示: 5个转基因纯合株系均为单拷贝; 纯合株系T_1Ab-10及其杂种的Bt蛋白含量最高; 人工接虫和自然感虫试验中, T_1Ab-10对二化螟、三化螟和稻纵卷叶螟均表现高度抗性; 喷药试验中, 阳性植株与阴性植株、对照植株的主要农艺性状均没有显著差异。这说明转cry1Ab基因纯合株系T_1Ab-10具商品化潜力, 可作基因聚合材料。     

根癌农杆菌介导法获得烟草转人骨形成蛋白—3成熟肽基因植株

利用冻融法将质粒pCAMBIA-hBMP-3m直接转入根癌农杆菌LBA4404,以烟草无菌苗叶盘为外植体,通过农杆菌介导法进行遗传转化,获得了在含25mg/L潮霉素（Hy-gromycin,Hyg）的筛选培养基上再生的抗性植株,经PCR检测呈阳性,初步鉴定并筛选整合了人骨形成蛋白－3成熟肽基因的转基因植株。     

CYP79F1基因对西兰花的遗传转化的研究

建立有效的遗传转化体系是基因工程手段改良植物性状的重要前提。本研究对西兰花遗传转化体系进行了初步研究,成功地建立了农杆菌介导的西兰花遗传转化体系。以7日龄无菌苗的下胚轴为外植体,预培养2~3 d后进行农杆菌的侵染,共培养后将外植体转移至无筛选压力的脱菌培养基中,暗培养7 d后置于含有10mg·L-1卡那霉素的筛选培养基中继续培养,待不定芽生长至2~3 cm时转入0.1 mg·L-1IBA的生根培养基中诱导不定根,最后进行驯化和移栽,PCR分子鉴定及外源基因表达水平检测结果证明,我们已经成功地将芥子油苷合成关键酶基因CYP79F1基因转入西兰花商业品种"青秀"中,利用HPLC检测转基因植株中短链脂肪族芥子油苷的含量显著增加。     

甜蛋白基因MBLII对番茄的遗传转化

以5～7d龄的 丽春 番茄无菌苗子叶作为外植体,研究了子叶外植体对抗生素卡那霉素的敏感性,抗生素卡那霉素对番茄筛选的适宜浓度为70mg/L.通过根癌农杆菌 Agrobacteriumtumefaciens 介导,成功地进行了马槟榔甜蛋白基因MBLII对番茄的遗传转化,获得转化番茄抗性植株,组织化学法有阳性表现、PCR特异扩增及Southern杂交检测出现特异条带,表明MBLII基因已顺利整合到转基因番茄植株的基因组.     

石斛兰转ACS反义基因抗性原球茎及转化植株的筛选与鉴定

该试验就石斛兰转化ACS(1-氨基环丙烷-1-羧酸合成酶)反义基因的不同筛选方法和筛选处理对抗性原球茎筛选的影响,以及石斛兰转基因植株的再生与鉴定进行研究.结果表明:(1)石斛兰原球茎经带有gus报告基因和ACS反义基因的农杆菌LBA4404侵染共培养5d后除菌,采用逐渐提高选择压浓度的延迟筛选,并在低选择压浓度下切割而高选择压浓度下不切割的处理方式为抗性原球茎的最佳筛选途径,抗性原球茎获得率可达14.97％.(2)抗性原球茎繁殖时应逐渐降低选择压浓度,且在低选择压浓度下进行切割处理,繁殖倍数达到1.15倍,且原球茎生长势好.(3)抗性原球茎在1/2 MS+0.5 mg/L 6-BA培养基中的分化率达到73.85％;107株无根小苗培养于1/2 MS+1.0 mg/L NAA+50.0 mg/L Km(卡那霉素)+100.0 mg/L Cef(头孢霉素)培养基中进行生根培养,共获得了13株具有卡那霉素抗性的转化植株,转化效率达到12.15％.(4)转化植株经报告基因产物GUS组织化学检测和gus的PCR检测,证实带ACS反义基因的T-DNA已整合进石斛兰基因组中,且转基因植株在形态上与未转基因植株无明显差别,3株转基因植株移栽2个月后均已成活.     

抗虫转基因甘蓝及其后代的研究

通过农杆菌感染法将苏云金杆菌杀虫蛋白基因转移进了甘蓝的基因组,带子叶柄的子叶作为外植体与农杆菌共培养.发生在子叶柄基部的愈伤组织在含卡那霉素(Km)15～30mg/L的MS培养基上进行筛选,约5%外植体上的愈伤组织继续长大,当移到含Km和6-BA的分化培养基上时,愈伤组织分化出绿色的芽.将芽分离培养,约80%在加有Km的培养基上被诱导生了根.未转化的对照组织在筛选培养基上不能分化出正常的芽和根系,并且逐渐褐化死亡.小菜粉蝶的幼虫被饲喂以转基因植株的叶片,幼虫出现中毒症状,表现为发育受阻和死亡.约20%受试植物的DNA与苏云金杆菌杀虫蛋白基因探针杂交显阳性带.由转基因植株的种子长成的第2代甘蓝幼苗的卡那霉素抗性和植株的抗小菜粉蝶的活力均符合孟德尔单基因分离规律.     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

Time of detection of recessive genes: effects of system of mating and number of examined individuals

Summary Anthers were cultured from two sets of seven lines of hexaploid wheat (Triticum aestivum L.) with different cytoplasms, the euplasmic nucleus donors, Siete Cerros 66 and Penjamo 62, as well as their six alloplasmic lines derived from wild relative species of the genera Triticum and Aegilops. Significant cytoplasmic and nuclear effects but no cytoplasmic-nuclear interaction were found for embryogenic anther response, with the best performance of Penjamo 62 in Ae. kotschyi cytoplasm. Plant regeneration was not affected significantly by the cytoplasmic background of the lines cultured. The possible genetic implications of the observed cytoplasmic and nuclear influences on the in vitro haploid induction of wheat are discussed.