组织培养与普通小麦异源易位系选育

以中国春小麦品种中８６０１、澳大利亚栽培小麦品种Ｓｕｎｓｔａｒ、Ｍｉｌｌｅｗａ作母本,与抗大麦黄矮病毒病（ＢａｒｌｅｙＹｅｌｌｏｗＤｗａｒｆＶｉｒｕｓ,简称ＢＹＤＶ）的小麦－中间偃麦草异附加系Ｌ１杂交,取杂种的幼胚和幼穗作离体培养,获得大量再生植株。经酶联免疫吸附分析（ＥＬＩＳＡ）、限制性内切酶长度多态性分析（ＲＦＬＰ）、染色体数目的检查和田间抗性鉴定,在再生植株回交后代中获得了抗ＢＹＤＶ的杂合易位株,经自交和花药培养纯合,选育出Ｄ１０９１、Ｄ１０９４、Ｄ１６５８、ＴＣ５、ＴＣ６、ＴＣ７等一批抗性稳定或基本稳定的普通小麦选系。以白黑麦６Ｒ二体附加系为抗源,与严重感染白粉病的陕７８５９杂交,经幼胚培养在再生植株回交后代中筛选到白粉病免疫的普通小麦杂合易位系。研究结果表明：在小麦远缘杂交中,组织培养可以作为导入外源基因产生易位的手段之一加以利用。     

利用phlb突变体创造普通小麦—簇毛麦６ＶＳ端二体代换系

用中国春phlb突变体（C.S.phlbphlb)与普通小麦－簇毛麦６Ｖ（６Ａ）异代换系（Ｓub.6V)杂交,再用phlb突变体与Ｆ１回交,在高配对植株中筛选出一个６Ｖ染色体发生变化的植株,编号为ＬＶ０２,用染色体Ｃ－分带和荧光原位杂交技术,对ＬＶ０２株系的后代进一步鉴定,在ＢＣ１Ｆ２中筛选鉴定出１株编号为ＬＶ０２－０１的植株,该株含有４０条普通小麦染色体,１条簇毛麦６Ｖ染色体和１条６Ｖ短臂端着丝粒染色体,在ＬＶ０２－０１的分离后代中用同样技术鉴定出８株普通小麦－族毛麦６ＶＳ端二体代换系。     

利用荧光原位杂交技术检测导入普通小麦的大赖草染色质

利用以大赖草基因组ＤＮＡ为探针的荧光原位杂交技术对导入普通小麦的大赖草染色质进行检测。结果：９１）根尖体中花粉母细胞时期均可用于检测大赖草染色质。间期核中杂交信号点数与所含大赖草染色体数目之间的对应关系依染色体显强Ｃ－带末端数不同而不同;（２）利用荧光原位杂交,从普通小麦－大赖草单体异附加系的减数分裂前植株＾６０Ｃｏ－γ射线辐射后代中检测到一批大赖草端着丝粒染色体和小麦－大赖草染色体易位等结构变异     

小麦-冰草异源附加系的创建 IF_3、F_2BC_1、BC_4和BC_3F_1世代的细胞学

为了进一步研究冰草属的Ｐ染色体组在小麦背景下的遗传效应和试图建立一套小麦－冰草属异源附加系,对来自普通小麦品种Ｆｕｋｕｈｏ×冰草Ｚ５５９杂种的Ｆ３、Ｆ２ＢＣ１、ＢＣ４和ＢＣ３Ｆ１世代的２２２株进行了减数分裂行为观察。结果表明：（１）２ｎ染色体的分布范围为３９～５４;（２）冰草Ｚ５５９的Ｐ染色体组存在抑制小麦Ｐｈ效应的遗传系统,而且该系统可能只涉及不多于３条Ｐ染色体,但其对Ｐｈ基因的抑制效应是微弱的;（３）Ｐ染色体组存在控制染色体在减数分裂后期分离的基因,该基因可能只涉及不多于２条Ｐ染色体;（４）抑制小麦ｐｈ效应的基因和控制染色体在后期分离的基因可能位于不同的Ｐ染色体上;（５）已获得５个可能的小麦－冰草异源附加系。     

普通小麦—鹅观草异附加系的选育与鉴定初报

应用根尖细胞染色体计数、花粉母细胞减数分裂染色体配对构型分析、植株外形特征观察、染色体Ｃ－分带技术,在普通小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ Ｌ．）－鹅观草（Ｒｏｅｇｎｅｒｉａ ｋａｍ ｏｊｉＯｈｗ ｉ）的杂交及回交后代Ｆ５、ＢＣ１Ｆ３、ＢＣ１Ｆ４和ＢＣ２Ｆ４群体中选育并鉴定出３个二体异附加系Ｖ３９－１５－５、Ｖ３５－８－８ 和Ｖ５８－６－１１。对植株外形特征观察法和Ｃ－分带技术在普通小麦－鹅观草异附加系的选育和鉴定过程中的作用等问题进行了讨论     

导入小麦的外源染色体片段的准确鉴定及外源抗性基因的稳定性分析

用抗白粉病的普通小麦一簇毛麦６ＶＳ／６ＡＬ易位系与普通小麦品种扬麦５号、普通小麦一簇毛麦６Ｖ代换系和中国春６Ａ双端二体以及６Ｖ代换系和６Ａ双端二体配制了４个测交组合,分析了这４个杂交组合Ｆ１ＰＭＣ’ｓＭＩＣ－分带的减数分裂构型。在（６ＶＳ／６ＡＬ易位系×扬麦５号）和（６ＶＳ／６ＡＬ易位系×６Ｖ代换系）的Ｆ１ＰＭＣ’ｓＭＩ,分别观察到由易位染色体与６Ａ染色体和６Ｖ染色体配对形成的具有特定Ｃ－分带带型的棒状二价体,杂种中的棒状二价体数目高于各自亲本中的棒状二价体数。在（易位系×Ｃ．Ｓ．ｄ．ｄ．ｔ６Ａ）Ｆ１中,在８７．９％的ＰＭＣ中观察到由易位染色体长臂与６ＡＬ端体配对形成的异形二价体（ｔｌ”）。而在（６Ｖ代换系×Ｃ．Ｓ．ｄ．ｄ．ｔ６Ａ）Ｆ１中,９６．６８％的ＰＭＣ具有两个单价端体（ｔ’,ｔ’）。该结果进一步证实易位涉及簇毛麦染色体６ＶＳ和小麦染色体６ＡＬ,易位断点靠近着丝粒。在减数分裂中,易位染色体的正常配对和分离,保证了６ＶＳ上白粉病抗性基因Ｐｍ２１的正常传递,为这一新抗源在小麦育种中的应用奠定了细胞学基础。     

小麦与高冰草属间体细胞杂交获可育杂种植株

普通小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．２ｎ＝４２）“济南１７７”与紫外线照射的高冰草（长穗偃麦草Ａｇｒｏｐｙｒｏｎｅｌｏｎｇａｔｕｍ,２ｎ＝７０）原生质体在ＰＥＧ诱导下融合,获杂种再生植株。取杂种子房诱导产生愈伤组织并再生植株,经染色体和同工酶鉴定,它们仍保留杂种性质。其中两株移栽成活并结实,杂种性质也经表型、染色体、同工酶和ＲＡＰＤ分析得到证明。在Ｆ０和Ｆ１代植株根尖细胞中,均发现高频率地存在着染色体断片;从Ｆ２代花粉母细胞减数分裂的染色体数目及行为发现,杂种细胞染色体数目主要分布在１８Ⅱ～２２Ⅱ,染色体断片发生配对及分离,表明它们是小染色体（ｍｉｎｉｃｈｒｏｍｏｓｏｍｅｓ）。Ｆ１及Ｆ２代植株比亲本小麦（“济南１７７”）秆茎粗硬、生长健壮,穗大粒大,已经产生具有优良性状的Ｆ２代穗系     

一个小黑麦附加—双代换花粉株系（M16）的创制与鉴定

报道了一个遗传组成复杂的小麦／小黑麦杂种花粉株系Ｍ１６的创制,形态性状,遗传稳定性,染色体组成以及抗白粉病的鉴定结果,Ｍ１６植株形态为普通小麦型,染色体数目为２ｎ＝４４,具有很高的遗传稳定性,有丝分裂和减数分裂的染色体稳定性分别为９１．０５％和９３．２３％,接种鉴定表明Ｍ１６高抗小麦白粉病,原位杂交,Ｃ－分带和同工酶标记分析证明Ｍ１６有３对黑麦染色体,是一个４Ｒ附加,３Ｒ（３Ｄ）和６Ｒ（６Ｄ）代换     

美味猕猴桃原生质体再生植株细胞遗传学研究 Ⅰ．体细胞染色体数目的变化

研究了美味猕猴桃叶愈伤组织原生质体再生植株和母株（ＡｃｔｉｎｉｄｉａｄｅｌｉｃｉｏｓａｌｉｎｅＮｏ．２６）茎尖体细胞染色体数目。结果表明：母株２ｎ＝６ｘ＝１７４。所测２９株再生植株的茎尖体细胞染色体数目差异显著,多为非整倍体类型,占所测植株的７２．４％左右;体细胞染色体数目介于１４２－３１０条之间,其中２ｎ＝６ｘ＝１７４约占２０．７％,少于１７４条染色体的植株约占３１．０％,超过１７４条染色体的植株则占４８．３％左右。个别单株部分茎尖体细胞在有丝分裂后期出现染色体桥、断片和落后染色体等异常现象。并对以上现象进行了扼要的讨论。     

抗白粉病6R(6B)染色体异代换系的选育

以中７９０２（６Ｂ）单体为母本,Ｈｏｌｄｆａｓｔ－ＫｉｎｇⅡ６Ｒ二体异附加系为父本配制杂交组合,经白粉病抗性追踪和细胞学鉴定,选育出抗白粉病６Ｒ（６Ｂ）二体异代换系。其与普通小麦中８６０１以及与“中国春”６Ｂ重双端体的Ｆ１植株,在减数分裂中期Ⅰ染色体构型分别为２０″＋２′和２０″＋１′＋２ｔ′ ,从而,证实６Ｂ染色体被 ６Ｒ染色体代换。同时,对６Ｒ染色体上抗白粉病基因的进一步利用进行了讨论。     

水稻原生质体再生植株及后代的性状表现

获得了粳型水稻77-170品系原生质体再生植株(R_2)6个株系的206棵后代植株。对其中96株进行性状观察和细胞学染色体鉴定,发现再生植株子代在株高、剑叶和主穗长、单株有效穗数、每穗粒数、育性、生育期等性状上都产生了变异,除株高外其他性状在第2代遗传上不稳定,染色体倍性稳定(2_n=24)。对56株再生植株子代作酯酶、过氧化物酶同工酶测定,其酶谱谱带与对照实生株相似。     

Somaclonal variant plants of wheat derived from mature embryo explants of three genotypes

Somaclones regenerated from three wheat (Triticum aestivum L.) cultivars, Glennson, Pavon and PAK-16171 were evaluated for variation in agronomic and morphological characters. Calli were initiated from germinating seeds on Linsmaier and Skoog (LS) medium plus 2 mg/l 2,4-dichlorophenoxyacetic acid, 2% sucrose and 1% agar. Calli were isolated and regenerated into whole plants on LS medium containing 0.1 mg/l indole - 3-acetic acid and 0.5 mg/l benzyladenine. Comparisons among the somaclones and their parents were made for plant height, spike length, number of grains per spike, and 100 grain weight. Significant variation was observed in these characters between the somaclones and parents. Genotypic differences were observed among the somaclones for many of these agronomic and morphological characters.     

起源于籼稻体细胞培养的联会消失变异的细胞遗传学研究

对表现不育的体细胞培养再生植株作减数分裂细胞遗传学分析发现,一株由IR54幼穗外植体起源的不育株(二倍体)为部分联会消失变异。其减数分裂早前期染色体配对正常,在终变期及中期Ⅰ观察到了数目不等的单价染色体,后期Ⅰ出现各种数目的落后染色体。由于减数分裂时染色体不平衡而导致该再生植株不育。     

Tissue culture in Haworthia

Summary Plants regenerated on two different media (NK and I) from the calluses of simple or cloned subcultures, which were originated from a single stock callus of Haworthia setata derived from its flower bud, were observed for eight characters, i.e., somatic chromosome number in root tips, growth vigor, leaf shape, leaf color, number of stomata per unit leaf area, esterase zymogram, chromosome association at meiotic metaphase I in pollen mother cells, and pollen fertility. From these regenerates plants with different characters from those of the parental plant were obtained. With regards to chromosomal aberrations, tetraploids, aneuploids, plants with a part of the chromosome segment deleted, with reciprocal and non-reciprocal translocations, or with paracentric inversions and those showing sub-chromatid aberrations at meiosis were obtained. The NK medium tended to regenerate more tetraploids and less plants carrying translocation than the I medium.Chromosome variabilities in somatic cells of the regenerates correlated with those of the calluses, from which they regenerated, while they did not correlate with either the meiotic irregularities (chromosome association at MI) or pollen fertility of the regenerates. From these facts, it was concluded that a rather large number of callus cells participate in the regeneration of an individual plant, although, however, only a few limited types of the cells form its germ line.Polyploidy affected growth vigor, leaf shape, stomata number and chromosome association at MI, but its effects were not detected on other characters. Chromosomal aberrations at the diploid level produced no clear changes in the regenerate's phenotype except in meiotic chromosome configuration and pollen fertility.Most chromosomal variants obtained in the present study are already reported in plants collected from wild populations, but plants with the deletion of a whole chromosome (karyotype 7L+6S) or chromosome segment (7L+1M+6S and 14L+2M+12S) have never been reported: this fact suggests that tissue culture is a powerful tool for producing plants with novel karyotypes.Contribution from the Laboratory of Genetics, Faculty of Agriculture, Kyoto University, Japan, No. 436     

应用基因组原位杂交鉴定蓝粒小麦及其诱变后代

应用基因组原位杂交技术（GISH）对普通小麦（Triticum aestivumL.)和长穗偃麦草[Agropyron elongatum(Host)Beauv,2n=10x=70]杂交后选育出的蓝粒小麦蓝-58及其诱变后代的染色体组成进行了鉴定。结果表明,GISH可方便地检测到小麦遗传背景中的长穗偃麦草染色体或易位的片段。如前人报道,蓝-58（2n=42)是一个具有2条长穗偃麦草4E染色体的异代换系（4E/4D）。LW004可能是一个具有两对相互易位染色体的纯合系,其田间表现磷高效特性,LW43-3-4为41条染色体的蓝单体（40W 1’4E）,种子颜色为浅蓝色,通过此法还检测出一些染色体结构发生很大变异的材料如4E的单端体（40W 1‘4E),种子颜色为浅蓝色,通过此法还检测出一些染色结构发生很大变异的材料如4E的单端体（40W 1‘t4E)以及组型为39W 1‘4E 1‘t4E的个体,此项研究结果更为直观地表明控制蓝粒体状的基因的确在来自长穗偃麦草的染色体上。同时说明有效的突变方法与灵活方便的检测手段的有机结合在染色体工程材料的创制和染色体工程育种中起着至关重要的作用。     

Induction of haploid and diploid plants though in vitro anther culture of haploid wheat (n=3x=21)

Summary Five haploid plants of wheat were used for anther culture. Embryos were formed and six plants were regenerated. Of these, two were haploid (n=3x=21) and two diploid (2n=6x=42). The two diploids derived from the anthers of the same haploid wheat plant gave seeds, but the fertility was reduced in one of them showing, abnormalities at meiosis.     

9个小麦品种(系)比较试验中的主要农艺性状

目的:通过对9个小麦品种(系)的比较试验中的主要农艺性状统计分析,筛选出比较适宜现阶段在贵阳地区乃至贵州省栽培的高产优质小麦品种(系)。方法:9个小麦品种(系)在试验地里随机区组排列,3次重复,记录小麦的全生育期,出苗数,田间生育期,株高,有效分蘖数,每株干重,穗长,穗粒重等,并在Excell上进行统计分析。结果:全生育期在200天左右,出苗数有3个超过100万/ha,最低只有44．8万/ha,株高在50 cm以上的有6个品种(系),3个在40 cm-50 cm之间,有效分蘖数400万/ha以上的有3个,最低的是199．1万/ha,每株干重最高是8．5 g,最低是5．7 g,穗长整齐度除98-22外都优于对照,经LSR法测验表明,贵农15,黔98353,黔麦15分别与夏繁29,98-28,贵单5号,黔98284-82,黔0110都达到差异显著水平,贵农15,黔98353,黔麦15分别与98-22达到差异极显著水平,其余的差异皆不明显。结论:可以初步得出贵农15,黔98353,黔麦15目前可以作为在贵阳地区推广种植的较优品种(系)。     

Stable progeny production of the amphidiploid resynthesized Brassica napus cv. Hanakkori, a newly bred vegetable

Resynthesized Brassica napus cv. Hanakkori (AACC, 2n?=?38) was produced by cross-hybridization between B. rapa (AA, 2n?=?20) and B. oleracea (CC, 2n?=?18) as a new vegetative crop. Many studies have provided evidences for the instability and close relationship between A and C genome in the resynthesized B. napus cultivars. In fact, seed produced to obtain progeny in Hanakkori had unstable morphological characters and generated many off-type plants. In this study, we investigated the pollen fertility, chromosome number, structure, and behavior linked to various Hanakkori phenotypes to define factors of unstable phenotypic expression in the progeny. Hanakkori phenotypes were categorized into five types. The results of pollen fertility, chromosome number, and fluorescence in situ hybridization analysis for somatic mitosis cells indicated that the off-type plants had lower pollen fertility, aberrant chromosome number, and structures with small chromosome fragments. Observation of chromosomes at meiosis showed that the meiotic division in off-type plants led to appreciably higher abnormalities than in on-type plants. However, polyvalent chromosomes were observed frequently in both on- and off-type plants in diplotene stage of meiosis. We assume that the unstable morphological characters in resynthesized progeny were the result of abnormal division in meiosis. It results as important that the plants of normal phenotype, chromosome structure and minimized abnormal meiosis are selected to stabilize progeny.     

Expression of various gametic types in pollen plants regenerated from hybrids between Triticum-Agropyron and wheat

Summary Studies of the chromosomal composition of pollen plants regenerated from the F₁ of hybrids produced from Triticum-Agropyron intermediate type and common wheat demonstrated that various gametic types of the F₁ could be fully expressed at the whole plant level via anther culture. The observed frequency of each of the eight types of pollen plants (based on their chromosome numbers) was in good agreement with the theoretical probabilities as shown by X² analysis. Comparative studies of the chromosome composition of somatic cells and pollen mother cells (PMC's) of selected pollen plants permitted classification of the plants into four distinct classes. The majority of these regenerated pollen plants had identical chromosome numbers in both root tip cells and PMC's. An alien disomic addition line, which was cytologically stable for two generations, was obtained directly from anther culture. Moreover, the addition line exhibits resistance to stripe rust disease, a trait which is conferred by the Agropyron chromosome. We suggest that anther culture techniques provide a unique and expeditious route for the introduction of alien genes or chromosomes into wheat cultivars.     

应用原位杂交技术对小麦—黑麦代换系间杂交的细胞遗传研究

应用原位杂交技术结合染色体组型分析方法,对两个小麦-黑麦异源双代换系5R/5A和6R/6A杂交后代的遗传进行了研究,探讨同祖染色体配对的可能性并获得小麦-黑麦易位系。实验中对杂种F1代植株减数分裂各时期的花粉母细胞染色体行为进行分析,结果发现有22.91%的花粉母细胞中黑麦染色体与小麦染色体发生同祖配对。F2代通过C-分带、原位杂交鉴定,在45株中检测到9株易位,易位频率为20%,是目前报道易位频率最高的。染色体易位有的来源于同祖配对交换,有的来源于单价体错分裂或断裂的重建。