苏云金芽孢杆菌Bt-3701与巨大芽孢杆菌Bm-107种间原生质体融合

具有杀虫活性的苏云金芽孢杆菌(B. thuringensis subsp kurstaki)Bt-3701与具有解磷活性的巨大芽孢杆菌(B. megaterium var. phosphaticum)Bm-107原生质体进行了融合。Bt-3701原生质体形成率为99.4％,再生率为21.4％;Bm-107原生质体形成率为97.5％,再生率为23.8％。以PEG为助融剂进行原生质体融合,得到22个融合子,融合率达6.03×10~(-3)。融合子在双抗培养基(DR-CNB)上连续传代12次,得到4个稳定的融合于生物测定表明,融合子既具有一定的杀虫活性又具有一定的解磷活性。     

芽孢杆菌原生质体的形成,再生及种间融合的研究

原生质体融合技术不仅能使遗传基因高频率重组,而且可以集双亲优良遗传性状为一体.自Schaeffer等人成功地进行了微生物原生质体融合以来,这项技术便广为人们所接受.枯草芽孢杆菌分泌的抗菌蛋白能抑制多种植物病原菌的生长,苏云金芽孢杆菌生成的伴孢晶体蛋白可毒杀植物害虫.利用原生质体融合技术将这两种芽孢杆菌抑菌杀虫的遗传特性融为一体,选育出防治植物病虫害的新一代生防菌株成为可能,有关这方面的研究国内外尚未见报道.本文报道了抗菌蛋白产生菌TG26-10和晶体蛋白产生菌AS1.904-17原生质体的形成,再生及种间融合的影响因素,为进一步筛选目的融合子提供基础.     

影响枯草杆菌原生质体转化的因素

原生质体转化是将外源基因导入细菌的主要方法之一,其转化频率受制于多种因素。本研究以激BacillussubtilisDB104为宿主菌,以枯草杆菌的高表达型质粒pNQ122为外源DNA,研究了原生质体再生率、原生质体浓度和用于制备原生质体的细胞生长期对转化频率的影响,获得了在该系统中实现高频率转化的条件。该转化条件使外源基因在多个B.subtilis菌株中的转化成为可能,并使从B.subtilis中筛选中性蛋白酶基因获得成功。     

INTERSPECIFIC CROSSES IN KALMIA

Interspecific crosses among Kalmia species are reported for the first time. All possible crosses between K. angustifolia, K. cuneata, K. latifolia, K. hirsuta, K. polifolia var. rosmarinifolia and K. p. var. microphylla were attempted. Several crosses yielded no hybrids, many produced albino or yellow-green inviable seedlings, and six combinations produced at least a few seedlings with near-normal growth habit. Stylar inhibition of pollen tube growth prevented some species from hybridizing; reciprocal differences in crossability and pollen tube growth were noted. Kalmia species are highly heterozygous, but no evidence was found within a species for geographic variation of combining ability except for the two varieties of K. polifolia which had different crossing patterns with the other species. The anthocyaninless form of K. angustifolia, in contrast to the normal, has unique combining ability with K. latifolia. No genetic evidence was found to support the placement of K. hirsuta in another genus (Kalmiella) or to designate K. angustifolia var. caroliniana as a distinct species. Use of all the species to breed improved ornamentals is limited by genetic barriers and sterility of F₁ hybrids.     

枯草芽孢杆菌在琼脂平板上进行的自然遗传转化

本文对发生在琼脂平板上的枯草芽孢杆菌自然遗传转化进行了初步的研究。结果表明,在相同条件下,该菌在琼脂平板上的自然转化率明显高于传统液体转化,并且转化反应对ＤＮａｓｅ 的抗性增强,通常被认为不能建立感受态的ＬＢ 培养物当涂布到平板上后也很快具有了自然转化的能力,说明在固相物表面进行的转化过程与传统的液体法存在一定的差异。在琼脂平板上,也能观察到具不同遗传标记的菌株间进行的细胞间自然转化。     

电脉冲法转化苏云金芽孢杆菌BMB171的研究

研究了用电脉冲法转化苏云金芽孢杆菌受体菌ＢＭＢ１７１的优化条件以及由转化导入的几类ｃｒｙ基因在ＢＭＢ１７１中的表达效果。结果表明,采用ＳＧ溶液作电脉冲缓冲液,用１０．ｏｋＶ／ｃｍ的脉冲场强和１次电脉冲（４．６ｍｓ）以及采用对数前期（ＯＤ_６５０ｎｍ为０．２～０．３）收获的受体菌,可以达到最高转化频率,其中用ｐＨＴ３１０１电转化 ＢＭＢ１７１的最高频率达 ８×１０７转化子／μｇ ＤＮＡ。转化频率随质粒ｐＨＴ３１０１浓度的增加,在５４．６９ｐｇ／ｍＬ至３．５０μｇ／ｍＬ范围内     

INTERSPECIFIC INCOMPATIBILITY IN SOLANUM SPECIES

Pandey , K.K. (Crop Res. Div., D.S. & I.R., Lincoln, Christchurch, New Zealand.) Interspecific incompatibility in Solanum species. Amer. Jour. Bot. 49(8): 874–882. Illus. 1962.—A diallel cross involving 11 self-incompatible and 3 self-compatible species of Solanum was made to study the genetic basis of interspecific incompatibility. Interspecific incompatibility was not limited to crosses in which a self-compatible species was used as the male parent onto a self-incompatible species (unilateral incompatibility). A number of crosses between self-incompatible species were incompatible. In one cross, Q vernei X verrucosum, a self-compatible species was successful as a pollen parent with a self-incompatible species. Unlike other hybrids between self-compatible and self-incompatible species, which are self-incompatible, these F₁ hybrids were self-fertile, and cross-fertile among themselves and with both parents. The self-fertile S. polyadenium was cross-incompatible as a female as well as a male parent with all other species. It is suggested that the unilateral incompatibility is a property of the allele S_C which originated as a consequence of one kind of breakdown of the S_I gene; the S_C allele produces “bare” pollen growth substances which are inactivated in an incompatible style. It is proposed that the failure of the principle of unilateral interspecific incompatibility in solanaceous species may be due to the action of alleles at the second incompatibility locus revealed in certain Mexican species. It is assumed that the South American species are selected intraspecifically only for the action of S alleles but that in certain interspecific crosses and rarely in intraspecific crosses the alleles at the second locus may be expressed, thus interfering with the usual action of S alleles. The F₁ hybrids Q verrucosum (self-fertile) X simplicifolium (self-sterile) were self-incompatible at the tetraploid as well as the diploid level, and their cross-compatibility behavior was consistent with the expected activity of the S_C and S_I alleles of the 2 parents respectively.     

猕猴桃属种间体细胞杂种

利用PEG融合方法,分别进行了中华猕猴桃(Actinidia chinensis var.chinensis)(2n=2x=58)子叶愈伤组织来源的原生质体与美味猕猴桃(A.deliciosa var.deiciosa)(2n=6x=174)子叶愈伤组织原生质体、以及狗枣猕猴桃(A.kolomikta)(2n=2x=58)叶肉原生质体种间原生质体融合。结果表明:中华猕猴桃与美味猕猴桃融合的1个克隆和中华猕猴桃与狗枣猕猴桃融合的4个克隆的RAPD谱带分别具有双亲特异的DNA谱带;经流式细胞仪分析,前者细胞核倍性推测为8倍体,后者细胞核为3倍体、4倍体和5倍体。初步鉴定这5个克隆是猕猴桃属种间体细胞杂种。     

苏云金芽孢杆菌无晶体突变株的逐级升温筛选及其转化性能

逐级从４２ ℃到４４ ℃和４６ ℃升温培养、并用０－０５ ％ ＳＤＳ 处理苏云金芽孢杆菌ＹＢＴ１４６３ ,获得了一系列内生质粒被部分或完全消除的无晶体（Ｃｒｙ －） 突变株,对４ 种Ｃｒｙ － 突变株的转化性能及导入的外源质粒的稳定性进行了研究。用限量培养基和４２ ℃培养筛选到Ｃｒｙ － 突变株后,升温至４４ ℃,从Ｃｒｙ － 突变株得到内生质粒被进一步消除的突变株;然后升温至４６ ℃来培养其中突变株ＢＭＢ１７０ ,并用０－０５ ％ 的ＳＤＳ进行处理,最终筛选到１ 株无质粒突变株ＢＭＢ１７１ 。用ｐＨＴ３１０１ 、ｐＢＭＢ１７３６ 、ｐＢＴＬ１ 和ｐＨＶ１２４９ 等４ 种外源质粒进行的转化及稳定性研究表明,转化频率的大小及导入质粒的稳定性与用作受体菌的Ｃｒｙ － 突变株携有的内生质粒数之间呈现一定的相关性,Ｃｒｙ－ 突变株的转化频率显著高于出发菌株,其中ＢＭＢ１７１ 的转化频率最高达１０７ 转化子／μｇ ＤＮＡ,且所导入的外源质粒的稳定性也高于其它Ｃｒｙ － 突变株及出发菌株ＹＢＴ１４６３ 。     

太白红杉种内和种间竞争研究

采用逐步扩大范围的方法确定影响对象木 (Objectivetree) 的最佳竞争范围, 利用单木竞争指数的改进模型对太白红杉 (Larixchinensis) 种内和种间竞争强度进行了定量分析, 并讨论了不同竞争强度下太白红杉的形态变化。结果表明 :随对象木胸径的增大, 由于太白红杉种群自然稀疏过程中密度调节作用, 植株距离增加, 种内竞争强度降低 ;太白红杉主要分布于亚高山地段, 群落内其它物种较少, 个体普遍较小, 结果种间竞争相对较弱, 种内与种间竞争关系顺序为 :太白红杉 太白红杉 >巴山冷杉 (Abiesfargesii) 太白红杉 >牛皮桦 (Betulaplatyphylla) 太白红杉 >其它树种 太白红杉 ;竞争强度和对象木胸径的关系服从幂函数关系 (CI =AD-B), 当太白红杉胸径达到 35cm以上时, 竞争强度几乎没有变化, 所得的预测模型能很好地预测太白红杉种内和种间的竞争强度 ;不同竞争强度下, 太白红杉主茎各层的分枝角度、总分枝数、当年生枝条长、平均枝长和活枝数均表现出显著的差异。表明采用逐步扩大范围的方法能有效地确定竞争木范围, 较好地反应太白红杉种内和种间的竞争关系。同时, 太白红杉通过自身形态变化, 提高了对光的截获能力和对不同竞争强度的适应能力。     

新疆甘草属的种间杂交

试验以居群为单位的新疆甘草属７个种进行种间人工杂交。通过对种间杂交结实量（ＲＳ）和亲本种平均结实量（ＲＳｍ和ＲＳｆ）的分析比较,测出相应的杂交结实指数（ＩＳ）、初步了解种间杂交亲和性的大小,为甘草属植物系统与演化的研究提供参考依据。     

NATURAL INTERSPECIFIC HYBRIDIZATION IN EASTERN NORTH AMERICAN CLAYTONIA

The eastern North American spring ephemeral Claytonia virginica (Portulacaceae) is well known for its great variation in chromosome number. The origin and significance of this cytological diversity has been a source of some controversy over the last two decades; in particular it has been suggested that one major source of variation could have arisen by allopolyploidy following hybridization between C. virginica and the second eastern North American representative of the genus, C. caroliniana. However, there has been no rigorous demonstration of natural hybridization between these taxa, and attempts to document hybridity have been hampered by a paucity of distinguishing morphological characters. Nuclear ribosomal gene markers are, however, able to distinguish between the two species and provide a means of identifying hybrid plants. We have found a locality in central New York state where the two species cooccur; in this population, hybrid individuals with additive ribosomal gene patterns are found in a zone of overlap between the two parental species. These hybrids are morphologically intermediate and have reduced pollen stainabilities, and appear to be F₁ individuals.     

HETEROCARYOSIS-LIKE PHENOMENON IN BACILLUS SUBTILIS.

Ikeda, Y. (University of Tokyo, Tokyo, Japan) and T. Iijima. Heterocaryosis-like phenomenon in Bacillus subtilis. J. Bacteriol. 83:1025-1028. 1962.-When a mixture of methionine-dependent, histidine-dependent, streptomycin-resistant cells and tryptophan-dependent, streptomycin-sensitive cells of Bacillus subtilis K is plated on minimal medium supplemented with asparagine and streptomycin, tiny colonies consisting of both component cells and X-type cells appear at a frequency of about 10(-6). Because the X-type cell segregates persistently both components and X-type, even after spore formation, it is considered to be a heterocaryon or a syncaryon possessing two kinds of genomes in a common cytoplasm.     

CARBON DIOXIDE FIXATION IN BACILLUS ANTHRACIS

Eastin, Jerry D. (U.S. Army Chemical Corps, Frederick, Md.) and Curtis B. Thorne. Carbon dioxide fixation in Bacillus anthracis J. Bacteriol. 85:410-417. 1963.-Virulent strains of Bacillus anthracis require a concentration of CO(2) greater than that of the normal atmosphere (air) for the production of capsular material (glutamyl polypeptide); avirulent strains may produce no polypeptide or may produce polypeptide in air. Fixation of C(14)O(2) by each of the three types tested resulted in labeling of aspartic acid, glycine, glutamic acid, succinic acid, and an unidentified organic acid. C(14) was detected in aspartic acid after less than 30 sec of exposure of cells to C(14)O(2). Subsequent flushing of the cells with C(12)O(2) displaced C(14) from aspartic acid but not from the other labeled intermediates. Aspartic acid appears to be closely associated with the primary CO(2)-fixation product, and the data suggest a fairly direct carbon pathway from CO(2) to aspartic acid (oxaloacetic acid) to glutamic acid to glutamyl polypeptide.