香菇和凤尾菇原生质体融合初探

选用经高温灭活的香菇原养型双核原生质体作供体亲本,以带有脯氨酸（Prol)营养缺陷型标记的凤尾菇单核原生质体为受体亲本,对其原生质体融合情况进行研究。结果发现,两亲本原生质体的融合再生率为0．106％,部分融合子菌丝细胞多核,无锁状联合。对该融合子再进行原生质体分离、再生,得到了具有新的生理特性的菌株材料。表明这种单双核原生质体融合是实现食用菌属间远缘杂交育种的有益探索。     

香菇和凤尾菇原生质体融合初探

选用经高温灭活的香菇原养型双核原生质体作供体亲本 ,以带有脯氨酸( Prol)营养缺陷型标记的凤尾菇单核原生质体为受体亲本 ,对其原生质体融合情况进行研究。结果发现 ,两亲本原生质体的融合再生率为 0 .1 0 6%,部分融合子菌丝细胞多核 ,无锁状联合。对该融合子再进行原生质体分离、再生 ,得到了具有新的生理特性的菌株材料。表明这种单双核原生质体融合是实现食用菌属间远缘杂交育种的有益探索     

用原生质体融合法获得稻恶苗菌异核体

用紫外线照射稻恶苗菌(Glbberella fujikuroi)野生型分生孢子9～12 min,存活率为0.6%～23%,得到单标记营养缺陷型突变菌株5株。选出1144(ade~-)、1184(arg~-)和1175(try~-)3株,用亚硝基胍诱变处理,得到双重标记的营养缺陷型突变菌8株。将11442(ade~-,pba~-)和11844(arg~-,try~-)两菌株用原生质体融合技术使它们形成异核体。用2%蜗牛酶加2%纤维素酶以0.6mol/L NaCl 液作渗透压稳定剂制成混合酶液,裂解幼嫩菌丝1.5～3 h,获得2×10'个/g 菌丝的原生质体。经35%PEG 融合剂将等量原生质体融合,获得异核菌落230个。经过进行排除营养互养的实验;对4株异核体菌株进行子代分离的遗传学分析;发现其两种核型的比例在1:1.4～1:8.5之间;对4株异核体还进行酯酶同功酶的测定。证明这4株菌株是异核体。     

黑木耳与毛木耳原生质体融合菌株扇变现象的研究

以原生质体融合得到的异核体菌株为材料其扇变部位，示扇变部位及亲本菌株的菌落特征，菌丝生长特性，菌丝细胞中的核相，营养需求，同工酶酶谱及它们间的拮抗反应等进行综合分析鉴定试验。试验结果表明：扇变部位，未扇变部位及亲本菌株间生物学特征与特性，同工酶谱显示出明显的差异。     

菌核侧耳子实体分化过程中的解剖学研究

通过对菌核侧耳子实体分化过程及不同阶段形态解剖特征的观察，发现菌核侧耳子实体分化可分为瘤状原基、棒状体、典型子实体3个阶段。瘤状原基、棒状体横切片可分为皮层、内层2层；菌褶为等式子实层类型，菌髓中菌丝排列方式属于混淆至不规则型；菌肉菌丝可分为生殖菌丝、联络菌丝和骨架菌丝3种类型，其生殖菌丝中单菌丝细胞间的锁状联合明显，菌丝间的锁状联合只在菌盖中出现。孢子印白色，孢子表面光滑，为柱状椭圆形。     

菌棱侧耳子实体分化过程中的解剖学研究

通过对菌核侧耳子实体分化过程及不同阶段形态解剖特征的观察,发现菌核侧耳子实体分化可分为瘤状原基、棒状体、典型子实体3个阶段.瘤状原基、棒状体横切片可分为皮层、内层2层;菌褶为等式子实层类型,菌髓中菌丝排列方式属于混淆至不规则型;菌肉菌丝可分为生殖菌丝、联络菌丝和骨架菌丝3种类型,其生殖菌丝中单菌丝细胞间的锁状联合明显,菌丝间的锁状联合只在菌盖中出现.孢子印白色,孢子表面光滑,为柱状椭圆形.     

OWE—SOJ技术及核迁移试验在香菇交配型因子鉴定中的应用

由于没胡明确的形态差异,香菇的３种不亲和交配反应即Ａ＝Ｂ＝,Ａ＝Ｂ≠和Ａ≠Ｂ＝难以互相区别。本研究将ＯＷＥ－ＳＯＪ技术及核迁移试验用于香菇的交配型分析。结果ＯＷＥ－ＳＯＪ技术,可以观察到３种不同的菌落形态：由Ａ≠Ｂ≠配对形成的具锁状联合的绒毛状菌落,Ａ≠Ｂ＝配对形成的无锁状联合的栅栏型菌落及由Ａ＝Ｂ＝或Ａ＝Ｂ≠配对形成的无锁状联合的绒毛状菌落。通过一个附加的常规交配试验,Ａ＝Ｂ＝和Ａ＝Ｂ≠反应易于     

维生素对继代培养中蛹虫草菌落形态及子实体产量的影响

研究维生素对蛹虫草继代培养中菌落类型及子实体产量的影响,旨在探讨优型菌落子实体产量最高,延缓退化的维生素配方.采用正交试验,将蛹虫草优良菌株J8在不同维生素培养基上进行10次继代培养.结果表明:菌落在不同维生素培养基中继代培养后形成5种菌落类型,有子实体形成能力的菌落为Ⅰ、Ⅱ、Ⅲ型,其中Ⅰ型菌落产量最高,Ⅲ型居中,Ⅱ型最低;Ⅳ、Ⅴ型无子实体形成能力.子实体产量最高,且最稳定的Ⅰ型为优型菌落.有利于优型菌落子实体高产的维生素配方为(VB1)2(VB2)3(VBC)4(VB6)2(VPP)3,并且5种维生素均有利于提高优型菌落子实体产量,但只有VB2对其子实体产量影响差异不显著.优型菌落形态特征为菌丝深黄色,气生性中等,菌丝致密绒毛状,有同心纹,有角变.     

淡紫拟青霉种内异核现象的初步研究

研究了淡紫拟青霉的不同田间分离菌株单孢子后代的异核现象。结果表明,来源于福建、江苏、安徽的4个菌株(19,20,459,641)的单孢子培养后代,在PPDA+1%活性炭的培养基上进行配接试验,仅在19和459 2个菌株间能形成异核体,其频率为12.25%;在配接试验中,2个亲和菌落交界处长出致密的菌丝簇组成的实线,可推测为异核体。从来自菌丝簇线的每个单菌丝尖端培养物中分离出30个以上的单孢子,将其分别移接到PDA平板上,将来源于单孢子的菌落与2个亲本菌落进行比较,有3个菌丝尖端培养物(459-8×19-2,459-7×19-10,459-6×19-1)重现了两亲本类型或出现了新的菌落类型,异核现象得到证实。     

灵芝原生质体融合HH系列菌株的选育研究

以灵芝菌株韩芝和红芝为亲本,进行原生质体融合,获得125株再生菌株,其中有锁状联合的菌株103株。与亲本的拮抗结果显示,在与双亲均有拮抗的20株中,10株能正常形成子实体,但菌株间性状差异显著,其中HH55、HH58和HH87具有突出的农艺性状,具有进一步开发应用的价值。     

Hybridization of Burkitt lymphoblastoid cells

Burkitt lymphoblastoid cell lines have been fused to mouse and human cell lines with the use of inactivated Sendai virus. The heterokaryons have developed into somatic cell hybrids of both parental cell types. Chromosome analyses confirm that cells now growing in selective medium are hybrids. Initial observations of preparations of the hybrid cells reveal that 5'-iododeoxyuridine can induce continued synthesis of Epstein-Barr virus antigens by these hybrid cells.     

Genetic transfer in plants through interspecific protoplast fusion

Protoplasts of sexually incompatible species have been fused and in some combinations have given rise to somatic hybrid plants. Partial elimination of parental chromosomes from either species is common in such hybrids, but total chromosome loss has generally occurred only with phylogenetically unrelated pairings. Genetic function of one parent may be retained despite a complete loss of its chromosomes, suggesting that genetic introgression is possible in the absence of complete donor chromosomes. A model interspecific combination for such studies is the potato-tomato somatic hybrid for which numerous phenotypes and karyotypes are encountered at the outset, with a broader range observed in the second somatic generation.     

Biochemical and cytological evidence for triple hybrid cell line formed from fusion of three different cells

Hybrids between two parental lines of somatic cells have been observed in various genomic multiplicities by ourselves and others. Possible explanations for the higher multiplicities include the fusion of cells in a 1:1 ratio with one genomic set undergoing an asynchronous replication either before or after fusion or the fusion of two or more cells. We now provide evidence for multiple genomic hybrids arising from the fusion of more than two cells in a mixture of three different cell lines. This proof is based on unique chromosomal and biochemical markers characteristic of the three parental cell lines. The distinctive phenotypes of all three lines are expressed in a clonally derived hybrid. Thus, we conclude that long-term, proliferating somatic cell hybrids can arise from the fusion of three or more cells.     

染色体原位杂交技术与植物体细胞杂种遗传鉴定

染色体原位杂交技术近年发展较快，已广泛应用于植物遗传育种实践。在植物体细胞杂种遗传鉴定方面，染色体原位杂交技术也日益显示出其优势，可用于倍性变异的来源检测，非对称杂种的非对称程度确认，融合双亲的基因组互作，染色体行为分析，以及融合双亲染色体在体细胞杂种有性过程中的传递等。还探讨了染色体原位杂交技术在柑桔远缘体细胞杂种遗传鉴定中的应用前景。     

苏皖地区栗疫病菌营养体亲和性研究

对江苏、安徽两省9个县市的栗疫病病原菌菌株进行了营养体亲和性(VC)研究,结果表明,不素和类型可分为3种:Ⅰ型为强不亲和,抗衡区两侧形成明显分生孢子器带;Ⅱ型为弱不亲和,抗衡区内不形成分生孢子器;Ⅲ型为中等不亲和,只在抗衡区的某一侧形成分生孢子器带。可将219个菌株划分为131个营养体亲和组(VCG_s),从而推算出至少有8个VC基因控制菌株的营养体亲和性。根据两省、田间小区以及大病斑上菌株的VCG_s分析,我国板栗疫病菌的VCG_s比其他国家的复杂。本文还探讨了复杂的原因及利用低毒力菌株进行生物防治的可能性。     

Murine leukemia virus: restriction in fused permissive and nonpermissive cells

Cultures of human cells nonpermissive for mouse leukemia virus replication could not be induced to support virus replication by homologous fusion in the presence of Moloney leukemia virus. Human cells were also fused with permissive mouse cells, and the fate of the virus in heterokaryons was determined by a simultaneous autoradiography and fluorescent antibody technique. Heterokaryons containing the full chromosome complement of both cells were likewise nonpermissive for virus synthesis, but hybrids of human and mouse cells, which lacked up to half of the human chromosome complement, were permissive for virus synthesis. The results suggest that human cell genes can direct a repressive control over mouse leukemia virus replication.     

Human serum albumin phenotype activation in mouse hepatoma--human leukocyte cell hybrids

Murine hepatoma cells that secreted mouse serum albumin were fused with human leukocytes that did not produce albumin. The resulting hybrids secreted both mouse and human serum albumin, as demonstrated by immuno-electrophoretic techniques. The activation of the human genome suggests that mapping genes governing specialized functions in somatic cell hybrids may be accomplished by using adifferentiated human cells as a parental line.     

一种快速鉴定柑桔体细胞杂种对脚腐病抗性的方法

通过用菌丝块接种柑桔叶片法进行柑桔体细胞杂种抗病性鉴定,并与接种柑桔枝条法,菌丝块接种嫁接苗法比较,证明柑桔不同部位对脚腐病的抗性存在着相关性,可采用这种相关性接种柑桔叶片进行体细胞杂种的抗病性鉴定。     

水稻籼粳交后代的籼粳分化与杂种F_1结实率的相关分析

以珍汕97B/秀水13的22个F_7代株系为材料,在采用程氏指数法和SSR分子标记进行籼粳属性鉴定分析的基础上,分别与4个光温敏两系不育系配制88个杂种一代,剖析供试材料的籼粳遗传背景与其配组的F_1结实率的相关性。结果表明:22个后代品系籼粳分化明显,大多为籼型或偏籼型;程氏指数法与SSR分子标记对供试材料的籼粳分类结果基本相符。F_1的结实率与双亲遗传背景呈极显著负相关,逐步回归分析结果表明光温敏不育系的籼粳分化和双亲的粳稻成分指数差值是决定F_1结实率高低的重要因素,携有广亲和基因的育种群体在籼粳交杂种优势利用上具有重要应用价值。     

Establishment and Identification of Cytoplasmic Male Sterility in Brassica napus by Intergeneric Somatic Hybridization

Exploitation of novel cytoplasmic male sterility(CMS)is a main approach for widening the cytoplasmic genetic background of hybrid oilseed rape and avoiding epidemic risk in oilseed rape production.In this study,symmetric somatic hybrids between Brassicanapus var.Zhongshuang4 and Sinapis arvensis(Yeyou18)were produced by protoplast fusion.Two of the six established hybrids were male sterile showing trace or no pollen release upon flowering with non-or slightly extended stamens.Using Zhongshuang4 as a recurrent parent to pollinate the male sterile plants,the ratio of male sterile plants increased with the number of backcrosses.As early as in BC3 generation,most of the sterile families had nearly 100%sterile plants.Up to BC4 generation,the male sterility became stable and no fertility segregation was observed.All F1 progenies from tested crosses using restorer and maintainer lines of Polima CMS were 100%sterile,indicating that the established CMS by somatic hybridization is different from Polima CMS.The origin of the cytoplasm and potential use of this hovel CMS in oilseed rape breeding were discussed.Key wotds:Oilseed rape,Protoplast fusion,Cytoplasmic male sterility,Sinapis arvensis