烟草种间的体细胞杂交──Ⅰ．通过原生质体融合将波缘烟草部分核基因组转移给普通烟草

通过“供体──受体”原生质体融合获得５个“波缘烟草＋普通烟草”体细胞杂种细胞系（ＴＵ＿１至ＴＵ＿５）。对５个杂种细胞系的细胞学观察表明,其中２个系为多细胞融合产物,其平均染色体数目分别为１６７．２和１９８．４;３个系为二细胞融合产物,其平均染色体数目分别为５４．３７０．６和６２．８。杂种细胞系的一个明显特征是所有观察的中期细胞均存在数量不等的畸变染色体。对５个杂种细胞系再生植株叶片苹果酸脱氢酶和酯酶的同工酶分析结果表明,它们均为程度不同的不对称核杂种,即只含有供体亲本（波缘烟草）的部分核基因组。以小麦ｒＤＮＡ为探针对杂种细胞系基因组ＤＮＡ的Ｓｏｕｔｈｅｒｎ杂交结果表明,杂种中双亲的载有ｒＤＮＡ的染色体可能均有部分消除;此外,杂种中新ＤＮＡ片段的存在说明双亲ｒＤＮＡ可能发生了分子间重组。两个由多细胞融合产生的杂种细胞系（ＴＵ＿２和ＴＵ＿５）再生苗不易生根而且形态上异常。３个由二细胞融合产生的杂种细胞系（ＴＵ＿１、ＴＵ＿３和ＴＵ＿４）再生苗可生根长大,形态上类似受体亲本;对其根尖的细胞学观察表明,染色体数目在５１─６６之间,说明均为只含有少量供体亲本染色体的不对称核杂种。     

Y染色体异常29例分析

本文从1992例遗传咨询病例中收集29例Y染色体异常的病例,其中Y染色体数 目异常(47,XYY)2例;Y染色体结构异常8例:Y/Y易位1例、Yp+3例、de l(Y)3例、嵌合 体dic(Y)1例;Y染色体长度变异19例。对Y染色体这几种异常类型的遗传效应进行分析。 Abstract:Twenty nine cases of Y chromosome abnormalities were found in 1992 patients asking genetic counseling.Different kinds of Y chromosome abnormalitics were detected by G and banding techniques.These were 47,XYY(2 cascs);46,X,del(Y)(3 cascs);46,X,Yp+(3 cases);46,X,t(Y;Y)(1 case);45,X/46,X,dic(Y)(1 case) and length changes of Y chromosome(19 cases).The genetic effects of Y chromosome abnormalities have been analyzed in this report.     

通过“供体-受体”原生质体融合将裸燕麦部分核基因组转移给普通小麦(英文)

本文进行了小麦和裸燕麦悬浮细胞原生质体的电融合,并基于双亲失活(用IOA处理受体小麦原生质体,用γ-射线照射供体裸燕麦细胞系),获得可能的杂种愈伤组织。对7块愈伤组织进行了乙醇脱氢酶(Adh)同工酶筛选,发现5块表现出双亲特征酶带。对3个杂种细胞系进行5种同工酶分析,证实它们均为稳定的不对称体细胞核杂种细胞系;它们表现出小麦的完整谱带和裸燕麦的部分谱带。对2个杂种细胞系及亲本的核糖体DNA Southern分析结果表明只有一个杂种细胞系(HB 95)含有双亲的全部谱带。细胞学观察表明,2个杂种细胞系的染色体数目均显著高于双亲。从杂种细胞系HB 94中分化出叶原基等分化结构。Adh同工酶分析表明,这些分化结构具有和母体细胞系完全相同的杂种谱带。     

294对自然流产夫妇的染色体分析

通过对294自然流产夫妇外周血染色体的检查,探讨了习惯性流产与染色体异常的关系。结果表明,异常核型24例中,染色体数目异常1例,结构异常22例,多太变异1例。染色体异常发生率是受检夫妇的8.16%（24/294）,是收件人群的4.1%（24/588）,高于一般人群染色体异常频率（0.5%）的8倍。并且发现10例世界首报核型。     

Asymmetric somatic hybrids between Lycopersicon esculentum and irradiated Lycopersicon peruvianum

Summary Asymmetric somatic hybrids of Lycopersicon esculentum and Lycopersicon peruvianum were obtained by fusion of leaf protoplasts from both species after irradiation of protoplasts or leaf tissue of L. peruvianum with 50, 300, or 1,000 Gy of gamma-rays. These radiation doses were sufficient to abolish the growth of the L. peruvianum protoplasts. The hybrids were selected for their ability to regenerate plants; this regeneration capacity derived from L. peruvianum. All asymmetric hybrid plants were aneuploid. The ploidy level, the morphology, and the regeneration rate were analyzed in relation to the radiation dose applied to L. peruvianum. After a low dose (50 Gy), most hybrids had near-triploid chromosome numbers, whereas after a high dose (300 or 1,000 Gy), most hybrids had near-pentaploid numbers. The morphology of the asymmetric hybrids was intermediate between that of L. esculentum and symmetric somatic hybrids of both species (obtained without irradiation treatment), and approached the morphology of L. esculentum to a greater extent after a high dose of irradiation. The asymmetric hybrids regenerated more slowly than the symmetric hybrids and regeneration proceeded more slowly after a high dose than after a low dose of irradiation. The high-dose hybrids also grew more slowly, flowered less, and set fruits less than the low-dose hybrids. No seeds could be obtained from any asymmetric hybrid.     

Asymmetric somatic hybrids between Lycopersicon esculentum and irradiated Lycopersicon peruvianum

Summary Asymmetric somatic hybrids of Lycopersicon esculentum and Lycopersicon peruvianum were analysed for the retention of genes and alleles specific for L. peruvianum. The hybrids were obtained by fusion of protoplasts from L. esculentum with those of L. peruvianum (the donor), the latter having been irradiated before fusion with 50, 300 or 1,000 Gy of gamma-rays. The retention of three different types of genes or alleles was analysed. (1) The gene coding for kanamycin resistance, which is dominant and had been introduced in most of the L. peruvianum donor plants by transformation. It was present at one locus in 16 L. peruvianum donor plants and at two loci in one donor plant. (2) The genes coding for acid phosphatase, locus Aps-1, and glutamate oxaloacetate transaminase (GOT); different alleles of these genes are co-dominant and were detected by isozyme analysis. (3) Eighteen single gene morphological markers for which most of the L. esculentum genotypes used were homozygous recessive. Kanamycin resistance from donor plants with one locus was retained in about 50% of the asymmetric 30H-hybrids (the donor was irradiated with 300 Gy). L. peruvianum specific alleles of Aps-1 and GOT were present in at least 70% of the hybrids; the retention of donor alleles was lower in 30H- than in 5H-hybrids (donor irradiated with 50 Gy). On average, 73% of the L. peruvianum-specific alleles (one or both) of the morphological markers were detected in the 30H-hybrids. Several of the L. esculentum genotypes used were homozygous recessive for two morphological markers on the same chromosome; in 43% of the 30H-hybrids derived from them, only one of these markers was complemented by the L. peruvianum allele. This is an indication of frequent breakage of the L. peruvianum chromosomes. Several hybrid calli regenerated genotypically different shoots. On the whole, this analyses confirms the conclusion drawn from the cytogenetic and morphological analysis of these asymmetric hybrids, namely that irradiation prior to fusion eliminates the L. peruvianum genome to only a limited extent.     

3种烟草基因组SSR位点信息分析和标记开发

利用生物信息学方法对绒毛状烟草、林烟草和本塞姆氏烟草3份烟草野生种基因组数据中的SSR位点信息进行了分析。结果表明,在全长分别为2.14Gb、2.44Gb和2.59Gb的绒毛状烟草、林烟草和本塞姆氏烟草基因组中分别获得153 357个、196 493个和278 784个SSR位点,平均相隔13.94kb、12.42kb和9.31kb出现一个SSR。在SSR位点分布区域上,绝大部分的SSR位点分布在内含子和UTR(尤其是5′-UTR)区域;在SSR基序类型上,主要集中在二、三碱基基序且二者占总SSR位点数目的80%以上,并以二碱基基序类型丰度最高;在SSR基序结构上,基因组中出现频率及数量最高的是含有A(T)n的基序结构;在SSR基序的重复次数上,除单碱基基序类型外,重复次数多在3~10次之间。利用分别属于5个不同烟草组的8份烟草材料验证所合成的300对引物,所有合成的引物均能扩增获得目标片段,其中有80对引物存在扩增多态性。表明来源于绒毛状烟草、林烟草和本塞姆氏烟草基因组的SSR标记在亲缘关系相对较近的烟草种间具有高度保守性和通用性,基于此3份烟草野生种基因组数据开发SSR引物用于后续的相关遗传研究具有可行性。     

GISH,AFLP and PCR-RFLP analysis of an intergeneric somatic hybrid combining Goutou sour orange and <Emphasis Type="Italic">Poncirus trifoliata</Emphasis>

Intergeneric somatic hybrids combining Goutou sour orange (Citrus aurantium L.) with trifoliate orange [Poncirus trifoliata (L.) Raf] were produced by electrofusion and their genetic inheritance analyzed by amplified fragment length polymorphism (AFLP), genomic in situ hybridization (GISH), and PCR-restriction fragment length polymorphism (PCR-RFLP). Sixteen mini-calluses were obtained after 20 days of culture; they all developed into embryoids on EME500 medium. Following several subcultures on shoot induction medium for a total culture period of 6 months, shoots regenerated. The plants grew vigorously with a well-developed root system and exhibited the trifoliate leaf character of P. trifoliata. Ploidy analysis verified that all of the regenerates were tetraploids (2n=4x=36) as expected. GISH analysis confirmed that 18 chromosomes came from trifoliate orange and the remaining 18 from Goutou sour orange, as with most symmetric somatic hybrid plants; moreover, chromosome translocations were also observed in one plant. AFLP analysis of 16 regenerates and their fusion parents indicated that all of the somatic hybrids except one were genetically uniform. Analysis of the somatic hybrid cytoplasmic genomes with universal primers revealed that their chloroplast DNA (cpDNA) banding patterns were identical to those of the mesophyll parent trifoliate orange, while their mitochondria (mt) genomes were of the callus parent sour orange. The potential of GISH in Citrus somatic hybrid analysis is discussed.The first two authors contributed equally to this paper.     

Single-Pair Fusion of Various Combinations Between Female Gametoplasts and Other Protoplasts in Nicotiana tabacum

This paper reports an enzymatic maceration-osmotic shock method for isolation of tobacco embryo sac and its component cell protoplasts, and also a new method for fusion between single pairs of selected mesophyll protoplasts using polyethylene glycol (PEG) as on inducing agent. An integration of these two methods has led to the successful fusion of female gametoplasts with other kinds of protoplasts. The female gametoplasts described here, in a broad sense, include the egg cell (E), central cell (C) and synergid (S). One of the female gametoplasts was selected and fused with another female, male (generative cell, G) or somatic (mesophyll, M)protoplast. Various combinations were involved: E+S, E+C, E +G, E+M, C+C, C+S, C+G, C+M, S+S, S+G, S+M, etc. Briefly, the authors were able to choose any desired combination to realize single-pair fusion by the new PEG method. For the purpose of culturing such fusion products that were limited in number, the authors had done some preliminary experimets using mesophyll protoplasts as feeder cells. Two methods were adopted: the microdrop culture, and the millicell culture with feeder cells. The mesophyll protoplasts were precultured for 2—3 days in large for population expansion before they were used as feeder cells. One or several protoplasts were cultured in a microdrop or a millicell and were induced formation of small cell clusters. This result indicated that the culture methods might also be suitable for culturing the products from fusion of female gametoplasts and other protoplasts in this plant species.     

小偃麦部分双二倍体及其异附加系异源染色体的GISH分析

应用ＴＩＳＨ对小偃麦部分双二倍体ＴＡＦ４６（２ｎ＝８ｘ＝５６）及其衍生的６个二体异附加系的中间偃麦草染色体组种类进行了分析、鉴定。以拟鹅冠草（Ｐｓ．ｓｔｒｉｇｏｓａ）ＤＮＡ为探针的分析结果表明,ＴＡＦ４６所含有的中间偃麦草染色体组为合成染色体组,即６条Ｓｔ组染色体和８条Ｅ组染色体。在其衍生的二体异附加系中,Ｌ４和Ｌ７含有Ｓｔ组染色体,Ｌ１、Ｌ２、Ｌ３、Ｌ５含有Ｅ组染色体。ＴＡＦ４６所含有的中间偃麦草染色体的部分同源群依次为ＩＥ（Ｌ３）、２Ｓｔ（Ｌ６）、３Ｅ（Ｌ２）、４Ｓｔ（Ｌ４）、５Ｅ（Ｌ５）、６Ｓｔ（Ｌ７）、７Ｅ（Ｌ１）。     

凤尾菇和桃红平菇种间原生质体电融合获杂种菌株

以有自然标记的双核风尾菇(Pleurotus sajor-caju)和桃红平菇(P．Rhodophyllus)为出发菌株,以它们携带营养缺陷型标记的单孢菌株为直接亲本,采用BAEKON 2 000基因转移仪进行侧耳属种间原生质体电融合,成功地获得若干株融合体,其中有一株编号为F57的融台体已培育出子实体。比较了融合体F57与亲本的形态、生理、生化和遗传等性状,结果证实,融合体F57是一株新的种间杂种菌株。