高产融合子Q4413的遗传学研究

本文通过对枯草芽孢杆菌BR151衍生株与北京棒杆菌1134衍生株的赖氨酸高产融合子Q4413的遗传型研究、生物量(菌体湿重、菌重、菌体体积和菌体密度)测定、DNA含量测定和GC比的研究,揭示了融合子在遗传学方面与双亲株之间的差异,进一步证实了该株确系稳定的重组子或新的融合子.     

原生质体融合选育赖氨酸高产菌种的研究

北京棒杆菌1134衍生株与枯草芽孢杆菌151衍生株的原生质体融合得到了一株可以甜菜糖蜜为原料,在摇床与小罐发酵均能稳定产赖氨酸6 50％以上的高产菌株——Q4413。Q441 3菌落形态类似于11 34株,而菌体形态区别于双亲,呈较短的杆状,无鞭毛,但有较稀疏的纤毛。生理生化研究表明：Q4413耐盐性、蔗糖发酵,石蕊牛奶试验产碱状况酷似于151株,而与1134株略有差异;运动性、糊精、柠檬酸盐利用,MR试验、酪素水解、明胶水解、石蕊牛奶还原,又酷似于1134株而区别于151株;04413的最适生长温度、pH值居双亲之间;Qq4413为原养型加三重抗性[Rifr,AECr Nalr];其细胞DNA含量与GC比测定结果均居双亲之中,而且生物量测定结果也均与双亲有一定的差异。     

细菌科间原生质体融合的研究

本文将赖氨酸产生菌——北京棒杆菌1134株进行标记,同时将枯草芽孢杆菌BR151菌株进一步标记后,进行科间原生质体融合的探讨,揭示了科间原生质体融合的可能性。经适宜的条件培养和预处理,再在适宜的条件下,酶解制备亲本株的原生质体,用PEG(MW6000)为聚合剂,经低温短时间处理,将1134衍生株与151衍生株的原生质体进行融合;并在改良的DM_3平皿上加入适量的细胞壁碎片(作引物)与适量的人血清白蛋白制剂,使其再生,结果棒杆菌(在酶解后2.5h.)与枯草芽孢杆菌(酶解后20min)的制备率均已达到99.98％以上,再生率分别达21.3％与90.8％以上。经适温培养48h后,从融合平皿上随机挑出若干个菌落。点种于CM母平皿上,再分别影印于10套选择培养基平皿上,进行鉴定,其融合率达到了3.9×10~(-4)。从菌落形态和菌体形态上可大体分为近1134类型,近151类型和中间类型三种。经连续传代五代后,从融合子中筛选山了几株在摇床上以甜菜糖蜜为原料。产酸水平捉高近一倍的稳定高产融合子。其中Q4413株的产酸率平均达6.56％,10立升小罐发酵产酸率6.52％,糖酸转化率为40.25％,经进一步验证,该菌株的产酸能力和遗传性均是稳定的,为远缘工业微生物育种开辟了新的可行途径。     

利用原生质体融合技术选育防治植物病虫害的基因重组菌株

本试验选用抗菌蛋白产生菌枯草芽孢杆菌(B.subtilis) TG26和晶体蛋白产生菌苏云金芽孢杆菌(B.thuringiesis subsp,pacifeus) AS1.904的营养缺陷型衍生株,在聚乙二醇的诱导下进行原生质体种间融合,获得了表现双亲遗传性状的种间融合菌株。融合率为7.52×10~6,融合子经传代10次,稳定率为19.5%。融合菌株的菌落和细胞形态与亲本株明显不同。通过SDS-聚丙烯酰胺凝胶电泳检测,融合菌株表达了亲本的抗菌蛋白和毒素蛋白。抑菌杀虫试验表明,融合重组菌株具有抑制多种植物病原菌和毒杀鳞翅目幼虫的能力。     

香菇原生质体融合及融合子的鉴定

以香菇（Ｌｅｎｔｉｎｕｓｅｄｏｄｅｓ）种内不同株一对亲和的单核菌丝（７４０２〈２〉和９１０１〈１２〉）为亲本,以碘乙酰胺失活７４０２〈２〉作为筛选标记,经过原生质体制备、ＰＥＧ融合及融合子再生等步骤,选育得融合子。融合子与双亲无拮抗性,在菌丝形态,核数目及可溶性蛋白质图谱、酯酶同工酶谱和过氧化物酶谱上均与双亲单核菌丝及担孢子杂交子有区别。将Ｋｎｏｗｌｔｏｎ等于１９８４年建立的一种分离高频再生衍生株的方法首次运用至食用菌中,使原生质体再生率提高２－３倍,为融合操作提供了方便。     

利用菌体酯酶同工酶和可溶性蛋白SDS—PAGE鉴定链霉菌融合子

利用酯酶同工酶技术鉴定链霉菌融合子证实,融合子SP-7和SR-T19的酯酶同工酶谱表现为介于双亲的中间型,并互不相同。此外,融合子SR-7还出现1条双亲所没有的决要酶带。表明融合子在融合过程中发生了真正的遗传重组,而不是亲本遗传物质的简单加合。菌体可溶性蛋白SDS－PAGE显示,融合子SR-7和SR-T19与双亲具有总体上的相似性,且可分辨的谱带数目和强度都超过了双亲,同时相互之间又有明显的不同,支持了这2个融合子均是双亲重组的后代。     

平菇与香菇属间原生质体融合的研究

通过分离和出菇试验,获得了纯化的平菇(Pleurotus sapidus)和香菇(Lentinus edodes)单孢系。用溶壁酶去细胞壁制备成原生质体。以PEG为融合诱导剂,诱导两者原生质体融合。1988年获得了可以出菇的融合子。这些融合子形成的子实体,从形态、生长习性和菌伞的颜色特征上都与双亲有明显的差异。大部分氨基酸含量介于双亲之间。同功酶分析也显示出融合子呈现与双亲不同的酶带。融合子出现的上述新性状,可能是平菇与香菇的原生质体基因重组的结果。     

秸杆降解菌株抗药性标记的筛选

两株菌株原生质体融合技术中,为了将融合子与双亲本分离出来,要对其中的至少一个亲本进行标记。该文从降解秸杆的两种不同的真菌出发,在两株菌株原生质融合前进行抗药性标记的筛选,为两种菌株融合后融合子的筛选和分离提供依据。本实验采用10种药物,对双亲本进行抗药性初筛,然后对临界药剂用量筛选,通过对比和重复实验,镜检和酶活测定,获得两种对白腐真菌抗药而康宁木霉非抗性的药品,氟康唑和十一烯酸真菌清,剂量分别为400μg/mg和500μg/mg。     

林青链霉菌双亲灭活原生质体融合的研究

分别以紫外线、热灭活性林肯链霉菌947和9502原生质体,然后进行灭活双亲的原生质体融合,从16株融合子筛选到林青霉素高产株。用双亲的互补营养缺陷型对林肯链霉菌原生质体的制备、融合、再生的部分条件进行了研究。发现含0．4％Gly和34％蔗糖的SM培养基最适于实验菌株原生质体的制备、再生。聚乙二醇（PEG）分子量对原生质体融合影响不大,其在P缓冲液中的浓度却很重要。含50％PEG的P缓冲液最有利于原生质体融合。     

林肯链霉菌双亲灭活原生质体融合的研究

分别以紫外线、热灭活林肯链霉菌 94 7和 95 0 2原生质体 ,然后进行灭活双亲的原生质体融合 ,从 1 6株融合子筛选到林肯霉素高产株。用双亲的互补营养缺陷型对林肯链霉菌原生质体的制备、融合、再生的部分条件进行了研究。发现含 0 .4 %Gly和 34 %蔗糖的SM培养基最适于实验菌株原生质体的制备、再生。聚乙二醇 (PEG)分子量对原生质体融合影响不大 ,其在P缓冲液中的浓度却很重要。含 5 0 %PEG的P缓冲液最有利于原生质体融合     

Microbial and reducing agents catalyze the rearrangement of taxanes

5alpha, 7beta, 9alpha, 10beta, 13alpha-Pentahydroxy-4(20),11(12)-taxadiene derivative 1 was converted to two unprecedented 1(15-->11)abeo-taxanes and a taxane derivative with a C10-C11 double bond by Absidia coerula ATCC 10738a. A similar compound was obtained from treatment with zinc of a triacetoxy-4(20),11(12)-taxadiene derivative.     

Analysis of fertility in somatic hybrids of Nicotiana rustica and N. tabacum and progeny over two sexual generations

Summary Somatic hybrid plants, produced between Nicotiana rustica and N. tabacum by heterokaryon isolation and culture and also by mutant complementation, were examined regarding their ability to set seed. From a total of seventeen independent somatic hybrids, three were found to be partially self-fertile while the others did not set seed. Differences regarding the methods of hybrid selection, parental varieties and chloroplast composition of hybrids did not appear to be significant regarding the ability of plants to set seed. Much variation in fertility was observed in subsequent generations and by recurrent selection of the most fertile, over two generations, it was possible to increase the level of self-fertility in some of the progeny. One R2 derivative possessed approximately a tenfold higher level of self-fertility than it's somatic hybrid parent. The presence of genetic markers from both parents were observed in all progeny indicating their hybrid nature.     

Morphological asymmetry of the abdomen and behavioral laterality in atyid shrimps

Morphological asymmetry and behavioral laterality in vertebrate species have been intensively studied in recent years, while comparable invertebrate studies are rare. Here we demonstrate asymmetry in the curvature of the abdomen and laterality in evasive responses for two atyid shrimps, Limnocaridina latipes and Neocaridina denticulata. The frequency distributions of the angle of the abdominal curvature in both species were discretely bimodal, suggesting that the two populations are composed of both left- and right-type individuals. In N. denticulata, behavioral analysis using high-speed filming illustrated that the escape direction for each individual, evoked by a sudden non-lateralized stimulus, was correlated with its abdominal curvature: left- (right-) type shrimp jumped back-leftward (-rightward) significantly more than often. A crossing experiment with N. denticulata indicated that the trait frequency in the F1 generation from two left-type parents differed significantly from that of the F1 generation from two right-type parents, and that the trait frequency for the F1 generation from parents of different laterality types did not deviate from random. That is, offspring laterality type is affected by the lateralities of the parents, indicating that abdominal dimorphism in shrimp is genetically derived. These results suggest that shrimp have an innate laterality that controls their escape direction, which in turn may affect prey-predator interactions in the aquatic community.     

The advantage of multiple cultural parents in the cultural transmission of stories

Recent mathematical modeling of repeated cultural transmission has shown that the rate at which culture is lost (due to imperfect transmission) will crucially depend on whether individuals receive transmissions from many cultural parents or only from one. However, the modeling assumptions leading up to this conclusion have so far not been empirically assessed. Here we do this for the special case of transmission chains where each individual either receives the same story twice from one cultural parent (and retransmits twice to a cultural child) or receives possibly different versions of the story from two cultural parents (and then retransmits to two cultural children). For this case, we first developed a more general mathematical model of cultural retention that takes into account the possibility of dependence of error rates between transmissions. In this model, under quite plausible assumptions, chains with two cultural parents will have superior retention of culture. This prediction was then tested in two experiments using both written and oral modes of transmission. In both cases, superior retention of culture was found in chains with two cultural parents. Estimation of model parameters indicated that error rates were not identical and independent between transmissions; instead, a primacy effect was suggested, such that the first transmission tends to have higher fidelity than the second transmission.     

An unusually large 5q duplication in an adult female subject: spreading of inactivation and in vitro instability of the derivative Xp/5q chromosome

An unbalanced translocation resulting in an unusually large partial 5q trisomy (5q11-5qter) and partial Xp monosomy (Xp11-Xpter) is reported in a 24 yr old woman with phenotypic abnormalities including gonadal dysgenesis and mental retardation. The karyotypes of the parents and the brother were found normal. Peripheral blood stimulated lymphocytes and cutaneous fibroblasts of the proband exhibited constantly, after BrdU incorporation, selective inactivation of the derivative X;5 chromosome spreading to the 5q duplicate segment. A variety of numerical and structural changes involving the derivative chromosome were observed in about 10% of cells of the cultured lymphoblastoid line established from the subject's lymphocytes. The extended 5q duplication, according to the literature, is generally accompanied by a severe phenotype and by developmental failure; it is therefore believe that genetic inactivation of the 5q duplicated region permitted the proband's development to adult age, despite the profound chromosomal imbalance.     

Conformation and stability of the constant fragment of the immunoglobulin light chain containing an intramolecular mercury bridge

The constant fragment of the immunoglobulin light chain in which the intramolecular disulfide bond is reduced (reduced CL fragment) assumes a conformation very similar to that of the intact CL fragment and contains two sulfhydryl groups buried in the interior of the molecule [Goto, Y., & Hamaguchi, K. (1979) J. Biochem. (Tokyo) 86, 1433-1441]. In order to understand the role of the disulfide bond, a derivative in which the disulfide bond is replaced by an S-Hg-S bond was prepared and its conformation and stability were studied. The derivative was prepared by reacting the reduced CL fragment with mercuric chloride. Kinetic studies showed that the reaction is rate-limited by the unfolding process of the reduced CL fragment. The mercury derivative was as compact as the intact CL or reduced CL fragment, and a tryptophyl residue was found to be buried near the S-Hg-S bond in the interior of the protein molecule. Judging from the circular dichroic spectrum, however, the beta-structure characteristic of the immunoglobulin fold was disturbed. The stability of the derivative to guanidine hydrochloride was lower than that of the intact CL fragment, but the unfolding transition was reversible and cooperative. Decreased stability of the mercury derivative is due to its folded conformation being distorted by introduction of the S-Hg-S bond.     

Maize heterosis affects the structure and dynamics of indigenous rhizospheric auxins-producing Pseudomonas populations

A rhizobacterial population of 2430 Pseudomonas isolates, originating from one maize hybrid and from its parents, was screened for auxins production. Four hundred and twelve isolates were found to be auxin producers (aia+), and 27 of them were also part of a previously described PhlD+ sub-population. Interestingly, most part of the aia(+)-PhlD+ isolates came from the hybrid. This finding indicates that heterosis allows an increased colonisation by multi-beneficial PGPR strains. Furthermore, results on the abundance and genetic diversity of aia+ isolates gave evidence that maize root colonisation by aia+ Pseudomonas is an inherited trait regulated by heterosis. In fact, two times more aia+ isolates were obtained from the rhizosphere of the hybrid than from the rhizospheres of the parents, and an amplified rDNA restriction analysis showed that the hybrid increases the genetic diversity of aia+ populations when compared to its parents.     

Impact of depth of pedigree and inclusion of historical data on the estimation of additive variance and breeding values in a sugarcane breeding program

Sugarcane breeders in Australia combine data across four selection programs to obtain estimates of breeding value for parents. When these data are combined with full pedigree information back to founding parents, computing limitations mean it is not possible to obtain information on all parents. Family data from one sugarcane selection program were analysed using two different genetic models to investigate how different depths of pedigree and amount of data affect the reliability of estimating breeding value of sugarcane parents. These were the parental and animal models. Additive variance components and breeding values estimated from different amounts of information were compared for both models. The accuracy of estimating additive variance components and breeding values improved as more pedigree information and historical data were included in analyses. However, adding years of data had a much larger effect on the estimation of variance components of the population, and breeding values of the parents. To accurately estimate breeding values of all sugarcane parents, a minimum of three generations of pedigree and 5 years of historical data were required, while more information (four generations of pedigree and 7 years of historical data) was required when identifying top parents to be selected for future cross pollination.     

Evidence for the hybrid origin of Lathyrus plitmannii (Leguminosae)

ABSTRACT

The hypothesis that Lathyrus plitmannii may be of hybrid origin was tested on a morphological data set, derived from measurements of 50 herbarium specimens, by employing the computer program HYWIN. Results showed that all the specimens of L. plitmannii were putative hybrids and that the highest ranked pair of specimens that best matches the morphologies of its parents was L. gorgonei and L. pseudocicera. Examination of the distribution patterns of the three species shows that L. plitmannii is confined to regions common to both putative parents and that hybrids could be formed, assuming that genetic barriers were overcome. Additional supporting evidence includes overlaps in the edaphic requirements and the habitats of the three species. Lathyrus plitmannii is recognised as a distinct species that should be regarded as derivative of hybridisation between L. gorgonei and L. pseudocicera.