Somatic hybridization as a tool for tomato breeding

Protoplast fusion can be used to produce somatic hybrids of species that cannot be obtained by sexual hybridization. The possibility to introgress genes from Solanum species into the cultivated tomato species Lycopersicon esculentum, and to obtain novel cytoplasm-nucleus combinations (cybrids) was considered as an important strategy to extend the genetic variation available for tomato breeding. Somatic hybrids between L. esculentum and other Lycopersicon species, as well as between L. esculentum and Solanum or Nicotiana species, have been produced. Specific mutants, genotypes with antibiotic resistances, and metabolic inhibition by iodoacetate or iodoacetamide and irradiation were used for the selection of hybrids. In addition, the improvement of protoplast culture techniques and the use of the favourable tissue culture traits derived from species such as L. peruvianum, which have been introduced into tomato by classical breeding, allowed the efficient recovery of somatic hybrids. However, the occurrence of somatic incongruity in fusion combinations of L. esculentum and Solanum and even more in L. esculentum and Nicotiana, did not allow the production of true cybrids and/or fertile hybrids, indicating the importance of both cytoplasm-nucleus and nucleus-nucleus interactions in somatic incongruity. Another problem with fusions between distantly related species is the strongly reduced fertility of the hybrids and the very limited homoeologous recombination between chromosomes of the parental species. Partial genome transfer from donor to recipient through microprotoplast (+) protoplast fusion, and the production of monosomic or disomic chromosome addition lines, light overcome some of these problems. In symmetric somatic hybrids between L. esculentum and S. tuberosum the occurrence of limited somatic and meiotic recombination was demonstrated. Fertile progeny plants could be obtained, though at a low frequency, when embryo rescue was performed on a large scale after backcrossing hexaploid somatic tomato (+) potato hybrids with a tetraploid potato genotype. The potential value of genomic in situ hybridization (GISH) and RFLPs for the analysis of the genome/chromosome composition of the hybrids has been demonstrated for intergeneric somatic hybrids between Lycopersicon and Solanum.Abbreviations cpDNA chloroplast DNA - mtDNA mitochondrial DNA     

蓝粒小麦4Ag染色体的显微分离与鉴定

本研究采用显微分离技术从蓝粒小麦(Triticum aestivum L.(2n=6x=42)×Thinopyrum ponticum Liu &Wang(2n=10x=70))根尖细胞中期分裂相中分离出具有4Ag染色体形态特征的单条染色体,对分离后的细胞进行原位杂交(FISH),结果显示细胞中所剩的和显微分离到的单条染色体均为4Ag染色体.利用Sau3A接头介导的PCR方法对分离出的单条4Ag染色体进行体外扩增,扩增的DNA片段大小约为200～2 000 bp,主要集中在250～750 bp之间.以DIG-dUTP标记的蓝粒基因组DNA为探针,与该扩增产物进行Southern杂交,结果表明显微分离出的染色体得到了有效的扩增.利用该分离染色体的PCR扩增产物为探针,对蓝粒小麦进行原位杂交,证明显微分离的染色体体外扩增片段确实来源于4Ag染色体.本研究拓宽了染色体显微分离的范围,为构建4Ag染色体文库和克隆位于该染色体上的重要农艺性状基因提供了新途径.     

农大4号与DS-1欧李正、反交F1代果实品质的遗传变异分析

为探究欧李果实品质的遗传规律,本研究选用欧李农大4号与DS-1及其正、反交后代群体为试验材料,测定双亲及后代群体果实的平均单果重、核重、纵横径、果形指数、可溶性固形物含量、类黄酮含量、总酚含量及抗氧化能力(DPPH、FRAP、ABTS),对其后代遗传规律进行分析。结果表明,所测定的指标均呈现正态或偏正态分布,呈连续变异,为多基因控制的数量遗传性状。纵径、横径、果形指数和可溶性固形物含量的变异系数均小于20%,后代未出现广泛分离,选择潜力较小,其中果形指数的变异系数最小,选择潜力最小,这与大多数欧李果实呈现扁圆形的情况一致。平均单果重、核重、类黄酮含量、总酚含量和抗氧化能力的变异系数均超过20%,后代广泛分离,选择潜力较大,尤其以平均单果重的选择潜力最大。平均单果重、纵横径受父本影响较大,以大果为父本时更易选出大果后代。果形指数不受父母本影响。可溶性固形物含量和核重表现明显的衰退变异,为趋小性遗传。以高类黄酮、总酚含量和抗氧化能力的植株为母本时,后代更易选出高类黄酮、总酚含量和抗氧化能力的优株。相关性分析将表型特征与内在物质含量相联系,为初步判断物质含量奠定重要基础。主成分分析提取了包括清除ABTS自由基和类黄酮含量的第一主成分和包括纵横径的第二主成分。聚类分析筛选出了具有较高生物活性、较强抗氧化能力和果实较大等不同特征的优株,可根据育种目标加以重点利用。本研究为欧李育种提供了重要参考的意义。     

Grazing of a common species of soil protozoa (Acanthamoeba castellanii) affects rhizosphere bacterial community composition and root architecture of rice (Oryza sativa L.)

We performed a controlled experiment with rice seedlings (Oryza sativa L.) growing in Petri dishes on homogeneous nutrient agar containing a simple rhizosphere food web consisting of a diverse bacterial community and a common soil protozoa, Acanthamoeba castellanii, as bacterial grazer. Presence of amoebae increased bacterial activity and significantly changed the community composition and spatial distribution of bacteria in the rhizosphere. In particular, Betaproteobacteria did benefit from protozoan grazing. We hypothesize that the changes in bacterial community composition affected the root architecture of rice plants. These effects on root architecture affect a fundamental aspect of plant productivity. Root systems in presence of protozoa were characterized by high numbers of elongated (L-type) laterals, those laterals that are a prerequisite for the construction of branched root systems. This was in sharp contrast to root system development in absence of protozoa, where high numbers of lateral root primordia and short (S-type) laterals occurred which did not grow out of the rhizosphere region of the axile root. As a consequence of nutrient release from grazed bacteria and changes in root architecture, the nitrogen content of rice shoots increased by 45% in presence of protozoa. Our study illustrates that interactions over three trophic levels, i.e. between plants, bacteria and protozoa significantly modify root architecture and nutrient uptake by plants.     

高粱与旱稻属间杂交部分遗传物质导入的F_3及BC_1表型鉴定

以高粱(S.BICOLOR)为母本、旱稻(O.SATIVA)为父本进行属间杂交,F1出现双亲没有的短芒和黄色柱头新性状,并能在F2、F3及BC1稳定遗传。F3在株高、穗长、穗型、分蘖性等性状上仍疯狂分离,并出现父本白色籽粒单株。BC1在株高、分蘖二性状上接近父本,回交效应非常明显,T测定无差异,并出现多子房新性状。高梁与旱稻杂交及回交,能创造变异,产生新性状,对创造矮秆、优质、多子房新种质十分有效。然而,二者杂交后代主体性状仍似母本,只表现父本少数性状,属于部分遗传物质导入。     

基因芯片技术检测5种马病毒

通过分子克隆技术获得马疱疹病毒1型(Equineherpesvirus1,EHV1)、马动脉炎病毒(Equinearteritisvirus,EAV)、马流感病毒(Equineinfluenzavirus,EIV)、马传染性贫血病毒(Equineinfectiousanaemiavirus,EIAV)和东部马脑脊髓炎病毒(East-ernequineencephalomyelitisvirus,EEEV)等5种病毒各一段高度保守的特异性基因片段,用芯片点样仪逐点分配到处理过的玻片上,制备成检测芯片。提取样品中的RNA,进行反转录和荧光标记后滴加到芯片上进行特异性杂交,对杂交结果进行扫描检测和计算机软件分析。结果显示,制备的基因芯片可同时检测和鉴别上述5种病毒,可检测到阳性杂交信号的最高稀释度为10-6的病毒液,约25个病毒DNA拷贝,但其它病毒材料未见红色荧光信号,证明了本方法的特异性。在进口马的隔离检疫期间,采集马鼻肺炎、马动脉炎中和抗体阳性但病毒分离阴性马匹的白细胞悬液,分别在EHV1和EAV位点处可检测到阳性杂交信号。证明基因芯片技术不但快速、准确和敏感,而且可同时进行多种病毒的检测。     

优质弱筋小麦扬辐麦2号的选育

采用扬麦 1 5 8与突变品系 1 90 1 2杂交 ,再经辐射诱变选育而成的优质弱筋小麦新品种扬辐麦 2号 ,其品质符合国家弱筋小麦标准 ,表现高产稳产 ,抗逆性强 ,千粒重高 ,后期熟相好 ,适应长江中下游淮南麦区种植     

高粱与旱稻属间杂交部分遗传物质导入的F3及BC1表型鉴定

以高粱（S．bicolor）为母本、旱稻（O．sativa）为父本进行属问杂交，F1出现双亲没有的短芒和黄色柱头新性状，并能在F2、F3及BC 1 稳定遗传。F1在株高、穗长、穗型、分蘖性等性状上仍疯狂分离，并出现父本白色籽粒单株。BC1在株高、分蘖二性状上接近父本，回交效应非常明显，t测定无差异，并出现多子房新性状。高梁与旱稻杂交及回交，能创造变异，产生新性状，对创造矮秆、优质、多子房新种质十分有效。然而，二者杂交后代主体性状仍似母本，只表现父本少数性状。属于部分遗传物质导入。     

Genetic resource enhancement by isolation of diversegenotypes from seed progeny in predominantly sterile rose scentedgeranium Pelargonium graveolens

Four new accessions (BSP1-4) of rose scentedgeranium Pelargonium graveolens were derivedfrom the spontaneous half-sib seed progeny of mostly sterilepopulations of the cultivar accession Bipuli. The seed producingplants of Bipuli had been growing among the populations of cultivaraccessions Hemanti and Kunti in the temperate agroclimate ofKodaikanal, Tamilnadu in India. The four new accessions were comparedwith the cultivar accessions Bipuli, Hemanti and Kunti in fieldexperiments under subtropical agroclimate of Lucknow, Uttar Pradeshfor essential oil yield and quality related traits and in terms oftheir RAPD profiles. The crop yield and essential oil parameters andDNA profiles of the four new accessions and three cultivar accessionsallowed the conclusion that the accessions BSP-1, BSP-2 andBSP-3 were Bipuli × Kunti hybrids and BSP-4was a Bipuli × Hemanti hybrid. The accession BSP-4demonstrated hybrid vigour in several of the essential oil yieldrelated traits; on average basis it out yielded other accessions by afactor of about 2.4. The essential oil of BSP-4 had 89%rhodinol content in which citronellol to geraniol ratio was 1:1 andthe contents of isomenthone, menthone, 10-epi--eudesmol, 6,9-guaiadiene, decanoic acid andisodecanoic acid were relatively lower than in the oils of accessionsBipuli and/or Hemanti. The present study has shown a way for thegeneration of new genotypes in rose scented geranium in which thecultivars have been vegetatively maintained for decades and thegenetic resources are scanty.     

Frankia共生固氮基因的分离与克隆

利用ＤＮＡ－ＤＮＡ杂交方法，分离细枝木麻黄的Ｆｒａｎｋｉａ菌株Ｃｏ０１的ｎｉｆ克隆ｐＣｃ１ＧＸ，赤杨内生菌株Ａｔ４的ｎｉｆ克隆ｐＡｔ１ＧＸ及沙棘的ＦｒａｎｋｉａＨｒ１８的ｎｉｆ克隆ｐＨｒ１８ＧＸ、ｐＨｒ１１－③ＧＸ。用基因功能互补法，从Ｆｒａｎｋｉａ菌株Ａｔ４的基因文库中分离到二个可能可互补豌豆根瘤菌ｎｏｄ基因功能的克隆ｐＡｔ２ＧＸ、ｐＡｔ３ＧＸ，并制作了ｐＡｔ２ＧＸ、ｐＡｔ３ＧＸ的亚克隆，予进一步实验，获得充分的证据证明ｐＡｔ２ＧＸ、ｐＡｔ３ＧＸ是否带有Ｆｒａｎｋｉａ菌株Ａｔ４的ｎｏｄ基因．