白菜型油菜,芥菜型油菜与诸葛菜的属间杂交合子胚的组织培养

TissueCultureofZygoticEmbryosofIntergenericHybridizationAmongBrassicacampestris,B．junceaandOrychophragmusviolaceusWUYan－You,JIANGJiu－Yu（InstituteofGeochemstry,TheAcademyofScience,Guiyang550002）1植物名称白菜型油菜（Brassicacampestris）品种川油8号（2n=20）、芥菜型油菜（B.juncea）品种沪州四棱（2n=36）和诸葛菜（Orychophragmusviolaceus,2n=24）。2材料类别白菜型油菜和芥菜型油菜作母本,诸葛菜作父本,人工授粉22d“萝卜”角果中的未成熟杂种胚。3培养条件分化培养基为MS＋6-BA2mg／L（单…     

芥菜型油菜与白菜正反杂交的胚胎学研究

利用荧光技术对芥菜型油菜（Brassica juncea）与白菜（B．pekinesis）种间正反杂交后花粉萌发和花粉管生长过程进行了观察。结果显示：芥菜型油菜与白菜正交授粉后,花粉在柱头上能正常萌发,多数花粉管沿花柱到达胚珠完成受精,且受精方式有珠孔受精、合点受精和中部受精,少量花粉管生长不正常,出现花粉管顶端膨大扭曲,花粉管分支等异常现象;反交授粉后,花粉在柱头上萌发时柱头乳突细胞产生强烈胼胝质反应,影响花粉管生长,只有少量花粉管通过花柱到达胚珠完成受精。用石蜡切片技术观察了正反杂交后杂种的胚胎发育,正交杂种胚胎发育较早,胚和胚乳生长较正常,杂种胚一般均能发育至成熟;反交杂种胚发育至心型期便不能继续发育,胚乳也停滞在游离核阶段并最终败育。综合分析表明,芥菜型油菜与白菜正反杂交都存在一定程度的受精不亲和性。     

芥菜型油菜与白菜正反杂交的胚胎学研究

利用荧光技术对芥菜型油菜(Brassica juncea)与白菜(B. pekinesis)种间正反杂交后花粉萌发和花粉管生长过程进行了观察。结果显示: 芥菜型油菜与白菜正交授粉后, 花粉在柱头上能正常萌发, 多数花粉管沿花柱到达胚珠完成受精, 且受精方式有珠孔受精、合点受精和中部受精, 少量花粉管生长不正常, 出现花粉管顶端膨大扭曲, 花粉管分支等异常现象; 反交授粉后, 花粉在柱头上萌发时柱头乳突细胞产生强烈胼胝质反应, 影响花粉管生长, 只有少量花粉管通过花柱到达胚珠完成受精。用石蜡切片技术观察了正反杂交后杂种的胚胎发育, 正交杂种胚胎发育较早, 胚和胚乳生长较正常, 杂种胚一般均能发育至成熟; 反交杂种胚发育至心型期便不能继续发育, 胚乳也停滞在游离核阶段并最终败育。综合分析 表明, 芥菜型油菜与白菜正反杂交都存在一定程度的受精不亲和性。     

小麦Kr基因在小麦与玉米或鸭茅状摩擦禾杂交中的失活

用３７个小麦（Ｔｒｉｔｉｃｕｍａｅｓｔｉｖｕｍ）品种（系）为母本,分别与黑麦（Ｓｅｃａｌｅｃｅｒｅａｌｅ）、球茎大麦（Ｈｏｒｄｅｕｍｂｕｌｂｏｓｕｍ）、玉米（Ｚｅａｍａｙｓ）和鸭茅状摩擦禾（Ｔｒｉｐｓａｃｕｍｄａｃｔｙｌｏｉｄｅｓ）杂交,比较其亲和性,小麦和玉米或鸭茅状摩擦禾杂交比小麦与黑麦或球茎大麦杂交的亲和性显著提高。携带着显性Ｋｒ１和Ｋｒ２基因的小麦品种Ｈｏｐｅ与黑麦杂交,不能形成胚,而与玉米及鸭茅状摩擦禾杂交时,成胚率分别达１６．００％和３２．５０％。表明控制小麦与黑麦及球茎大麦杂交亲和性的Ｋｒ基因系统在小麦与玉米及小麦与鸭茅状摩擦禾属间杂交中失活。讨论了还存在有其它控制小麦属间杂交亲和性的遗传调控系统的可能性。     

白菜型油菜与蓝花子的属间杂交合子胚组织培养

TissueCultureofZygoticEmbryosofIntergenericHybridizationBetweenBrassicacampestrisandRaphanussativusvar．raphanistroidesWUYan-You,JIANGJiu-Yu（InstituteofGeochemstry,TheChimeseAcademyofScience,Guiyang550002）1植物名称白菜型油菜（Brassicacam－pestris）品种川油8号（2n=20）和蓝花子（Raphanussativusvar．raphanistroidesMakino,2n＝18）。2材料类别白菜型油菜作母本,蓝花子作父本,人工授粉22d“萝卜”角果中的未成熟杂种胚。3培养条件分化培养基为MS＋6－BA2mg／L（单位下同）＋NAA0．1;生根培…     

十字花科几种植物的胚培养研究

利用幼胚成功地培养出白菜型油菜、芥菜型油菜、甘蓝型油菜、埃塞俄比亚芥、诸葛菜的再生植株。通过研究表明：不同的植物在培养基中的反应不同。同一个材料,在不同时期,培养的效果也不同。白菜型油菜难形成愈伤组织,且愈伤组织也难以分化,但生根较易。其它几种芸苔属植物则易形成愈伤组织和分化。诸葛菜能够形成胚状体,胚状体极易萌发。除了白菜型油菜外,所有材料都表现出极强的再生能力。     

芥菜型油菜和白菜型油菜种间杂种遗传分析

种间杂交是一种拓宽栽培作物遗传基础和转移优良性状的重要手段,已经广泛地用于作物品质的改良。本研究通过芥菜型油菜（Brassica juncea L.）和白菜型油菜（Brassica rapa L.）种间杂交,将芥菜型油菜的有利性状转移到白菜型油菜中,创造新型白菜型油菜,以改良白菜型油菜的农艺性状、提高抗逆性和拓宽其遗传基础。研究结果表明：以芥菜型油菜作母本、白菜型油菜作父本的杂交组合较易获得杂交种子,杂种F1植株营养生长具有较明显的杂种优势,但花粉完全不育;以白菜型油菜回交获得的BC1植株间表型差异明显,平均花粉可染率为34.8%,介于 0~84%之间,群体自交不亲和;BC1F1和BC2群体变异广泛,出现自交亲和植株和黄籽植株,平均花粉可染率分别为79.7%和79.1%。     

高频率产生芸苔属非整倍体和纯合植株及基因组原位杂交分析

进行了芥菜型油菜(Brassica juncea (L.) Czern. &; Coss.)和埃塞俄比亚芥(B. carinata A. Braun)与新油料植物资源诸葛菜(Orychophrogmus violaceus (L.) O. E. Schulz)之间的属间杂交, 并对产生的后代植株进行了基因组原位杂交分析. 结果表明, 芥菜型油菜与诸葛菜的杂交后代由3类混倍体组成, 第一类的体细胞和花粉母细胞(PMC)主要含有芥菜型油菜的36条染色体和附加的诸葛菜染色体(2n = 36 ~ 44); 第二类(2n = 30 ~ 36)主要产生具有芥菜型油菜染色体的PMC(2n = 36)和1 ~ 4对芥菜型油菜染色体被诸葛菜染色体所代换的代换型PMC(2n = 36); 第三类(2n = 30 ~ 36)植株的PMC则只具有芥菜型油菜染色体(2n = 36). 而埃塞俄比亚芥与诸葛菜的杂交后代由两类混倍体(2n= 29 ~ 34)组成, 第一类的绝大多数PMC的染色体数目(2n= 34)及行为与母本埃塞俄比亚芥植株的一样, 只是部分PMC包含1 ~ 3对诸葛菜染色体, 而第二类的PMC只具有埃塞俄比亚芥的34条染色体. 从以上杂种的后代中获得了芸苔属亲本种的附加系、代换系、 亚倍体和纯合植株. 这些结果为在芸苔属与诸葛菜属间杂交中可能发生的亲本种染色体组分开现象提供了分子细胞遗传学证据.     

远缘杂交油菜核不育系的创建及其细胞学和形态学研究

在甘蓝型油菜与诸葛菜以及芥菜型油菜与诸葛菜属间杂交后代中分别发现1个和3个不育材料,经杂交和多代近交育成了相应的甘蓝型油菜不育系。通过对核不育系体细胞鉴定表明,所有新发现的不育系染色体数为38,均已恢复到甘蓝型油菜。这些不育系绝大部分花粉母细胞（PMC）在中期Ⅰ、后期Ⅰ和后期Ⅱ 3个时期染色体行为表现正常,但不同时期的PMC均会出现一定比例的异常现象,主要表现为染色体落后或染色体桥等。这些不育系属于单核败育型,不育株与可育株的花器形态差异明显,不育系还存在不同程度的死蕾等特点。通过对花器生长过程的研究,发现不育株雌蕊生长随雄蕊败育进程逐渐加快,而可育株雌蕊生长则存在两个生长缓慢阶段。此外,文章还讨论了这些不育系的应用前景。     

油菜×蔊菜远缘杂交创新油菜种质资源研究

本项研究重点是要探讨通过油菜与蔊菜远缘杂交,将蔊菜的某些优良性状转移到栽培油菜中的可行性.结果表明甘蓝型油菜与蔊菜进行远缘杂交有较好的亲和性.而且其杂交后代变异明显,出现了许多碎叶型植株和一些类似白菜型的植株,杂交后代对菌核病有较强的抗性,有许多具有优势的单株.油菜与蔊菜进行远缘杂交对于创新油菜种质资源,培育高产、高抗油菜新品种具有重要意义.     

High efficiency production and genomic<Emphasis Type="Italic">in situ</Emphasis> hybridization analysis of<Emphasis Type="Italic">Brassica</Emphasis> aneuploids and homozygous plants

Interspecific and intergeneric hybridizations have been widely used in plant genetics and breeding to construct stocks for genetic analysis and to introduce into crops the desirable traits and genes from their relatives. The intergeneric crosses between Brassica juncea (L.) Czern. & Coss., B. carinata A. Braun and Orychophragmus violaceus (L.) O. E. Schulz were made and the plants produced were subjected to genomic in situ hybridization analysis. The mixoploids from the cross with B. juncea were divided into three groups. The partially fertile mixoploids in the first group (2n = 36-42) mainly contained the somatic cells and pollen mother cells (PMCs) with the 36 chromosomes of B. juncea and additional chromosomes of O. violaceus. The mixoploids (2n = 30-36) in the second and third groups were morphologically quite similar to the mother plants B. juncea and showed nearly normal fertility. The plants in the second group produced the majority of PMCs (2n = 36) with their chromosomes paired and segregated normally, but 1-4 pairs of the O. violaceus chromosomes were included in some PMCs. The plants in the third group produced only PMCs with the 36 B. juncea chromosomes, which were paired and segregated normally. The mixoploids (2n = 29-34) from the cross with B. carinata produced the majority of PMCs (2n = 34) with normal chromosome pairing and segregation, but some plants had some PMCs with 1-3 pairs of chromosomes from O. violaceus and other plants had only PMCs with the B. carinata chromosomes. The Brassica homozygous plants and aneuploids with complete or partial chromosome complements of Brassica parents and various numbers of O. violaceus chromosomes were derived from these progeny plants. The results in this study provided the molecular cytogenetic evidence for the separation of parental genomes which was previously proposed to occur in the hybridizations of these two genera.     

High efficiency production and genomic in situ hybridization analysis of Brassica aneuploids and homozygous plants

Inplantbreedingandgeneticresearch,karyotypicallystablecrosseswhichproducehybridplantshavebeenextensivelyusedtointroduceintocropsthetargettraitsandgenesfromrelatedwildorcultivatedspeciesortoconstructstocksforgeneticanalysis(alienchromosomeadditions,substitutionsandtranslocations)[1—3].Uniparentalgenomeeliminationinkaryotypicallyunstablehybridshasbeenutilizedforhaploidproduction[2,4].Becausetheartificiallysynthesizedallopoly-ploidscannotbeusedascropsformanyreasons,onepurposeofwidehybridizations…     

十字花科10属种与油菜萝卜胞质不育系杂交的花粉萌发情况观察

对10属种十字花科植物与油菜萝卜胞质不育系杂交时花粉在柱头上粘合、萌发、花粉管伸长等情况进行观察。结果表明：（1）海甘蓝花粉粒粘合较难;（2）48 h内无瓣焊菜〖WTBX〗(Rorippa dubia)〖WTBX〗、毛果诸葛菜(Orychophragmus violaceus)、桂竹香(Cheiranthus cheiri)、海甘蓝(Crambe abyssinica)花粉管的伸长受阻于花粉萌发启动之时,花粉壁内形成胼胝质塞;播娘蒿、紫罗兰、荠菜花粉管伸长但未进入乳突细胞;芝麻菜花粉管进入乳突细胞而未进入柱头,‘浠水白’(Brassica campestris)、蓝花子有花粉管进入柱头及花柱而未进入胚囊。     

Genome doubling and chromosome elimination with fragment recombination leading to the formation of Brassica rapa-type plants with genomic alterations in crosses with Orychophragmus violaceus.

In distant hybridization of plants, nonclassical hybrids with unexpected chromosome complements, chromosome elimination, and genetic introgression have been well documented. We obtained intergeneric hybrids between Brassica rapa, B. rapa var. chinensis, and another cruciferous species, Orychophragmus violaceus, following embryo rescue. Hybrids mainly displayed phenotypes of B. rapa, although certain O. violaceus or novel characteristics also appeared. Variable numbers of chromosomes were observed in somatic cells in the roots of plantlets on medium and in ovaries and pollen mother cells (PMCs). However, higher numbers were recorded in the roots. GISH revealed that the majority of ovary cells and PMCs contained 20 chromosomes of B. rapa with or without individual O. violaceus chromosomes or fragments added or introgressed. AFLP analysis showed that fragments deleted from the B. rapa genome were much more frequent than novel and O. violaceus fragments. The mechanisms involved genome doubling and successive elimination of O. violaceus chromosomes accompanied by fragment recombination and introgression, producing B. rapa-type plants with modified genetic constitutions and phenotypes.     

甘蓝型油菜与诸葛菜属间杂种的减数分裂观察

在甘蓝型油莱与诸葛菜属间杂种细胞减数分裂中,观察到两种类型的染色体分离方式。第一种,细胞内的３１条染色体呈两组分离,其中１９条染色体至细胞一极,１２条染色体至另一极;且在两极的染色体进行分裂,产生４个核,两个具有１９条染色体,另两个具有１２条染色体。故杂种细胞通过这种染色体分离方式,可同时产生具有１９与１２条染色体的两类配子。杂种细胞内发生的这种染色体行为,极有可能与来自两个亲本种的染色体组相关;即在细胞两极的１９与１２条染色体分别来自甘蓝型油菜与诸葛菜。第二种,在后期Ｉ至末期Ｉ,花粉母细胞内以６个二价体形式出现的１２条染色体被落后在赤道板附近,而被排斥在末期核之外。这些落后二价体很有可能来自诸葛菜。因此,在杂种细胞减数分裂中,来自两个亲本种的染色体组被分开。文中还对染色体组分开的遗传意义等进行了讨论。     

Cytogenetic and molecular characterization of intergeneric hybrids between Brassica napus and Orychophragmus violaceus.

Twenty-two intergeneric hybrids from a cross between Brassica napus (AACC, 2n = 38) cultivar Oro and the ornamental crucifer Orychophragmus violaceus (OO, 2n = 24) were produced without embryo rescue. The plants were classified into three groups based on morphological and cytological observations and RAPD banding patterns. Plants of Group I had morphological traits of both parents and 2n = 29 chromosomes. In these plants, 62.1% of the pollen mother cells (PMCs) had the pairing configuration 1 III + 9 II + 8 I; the remaining PMCs had 10 II + 9 I. The plants possessed 97.6-98.8% B. napus specific and 9.2-11.7% O. violaceus specific RAPD fragments. Plants of Group II exhibited novel morphological traits and possessed 2n = 35, 36, or 37 chromosomes. Plants of Group III were morphologically similar to B. napus and possessed 2n = 19, 37, 38, or 39 chromosomes. Plants of Group II and Group III had 94.1-99.4% B. napus specific RAPD fragments and no O. violaceus specific RAPD fragments. Chromosome fragments were observed in PMCs of most of the F1 plants in all groups. Based on the cytological results and RAPD analysis, it is suggested that genome doubling and chromosome elimination occurred in the intergeneric hybrids of B. napus x O. violaceus.     

Parental genome separation and elimination of cells and chromosomes revealed by AFLP and GISH analyses in a Brassica carinata x Orychophragmus violaceus cross

BACKGROUND AND AIMS: The phenomenon of parental genome separation during the mitotic divisions of hybrid cells was proposed to occur under genetic control in intergeneric hybrids between cultivated Brassica species and Orychophragmus violaceus (2n = 24). To elucidate further the cytological and molecular mechanisms behind parental genome separation, Brassica carinata (2n = 34) x O. violaceus hybrids were resynthesized and their chromosome/genomic complements analysed. METHODS: F(1) hybrids of the cross were obtained following embryo rescue, and were investigated for their cytological behaviour and subjected to genomic in situ hybridization (GISH) and amplified fragment length polymorphism (AFLP) to determine the contribution of parental genomes. KEY RESULTS: All the F(1) plants with high fertility closely resembled B. carinata in morphological attributes. These were mixoploids with 2n chromosome numbers ranging from 17 to 35; however, 34, the same number as in B. carinata, was the most frequent number of chromosomes in ovary and pollen mother cells (PMCs). GISH clearly identified 16 chromosomes of B. nigra in ovary cells and PMCs with 2n = 34 and 35. However, no O. violaceus chromosome was detected, indicating the presence of the intact B. carinata genome and elimination of the entire O. violaceus genome. However, some AFLP bands specific for O. violaceus and novel for the two parents were detected in the leaves. Cells with fewer than 34 chromosomes had lost some B. oleracea chromosomes. F(2) plants were predominantly like B. carinata, but some contained O. violaceus characters. CONCLUSIONS: The cytological mechanism for the results involves complete and partial genome separation at mitosis in embryos of F(1) plants followed by chromosome doubling, elimination of cells with O. violaceus chromosomes and some introgression of O. violaceus genetic information.     

甘蓝型油菜与诸葛菜属间杂种无性系的变异研究

从形态及细胞学两方面对甘蓝型油菜与诸葛菜属间杂种无性系的变异进行了研究。经过长期继代培养后,杂种在形态上越来越偏向母本甘蓝型油菜,而表现出较少的父本诸葛菜性状,但表现出对等双分枝的新性状。经细胞学观察表明,杂种体内杂种细胞比例下降,而甘蓝型油菜细胞比例大幅度上升并远高于核质杂种细胞（具有油菜细胞质与诸葛菜细胞核）的比例,以至较多的甘蓝型油菜染色体组被遗传下去。在该杂种继代培养中还观察到杂种细胞内的染色体消除及体细胞配对现象。     

Alteration of chromosome behavior and synchronization of parental chromosomes after successive generations in Brassica napus x Orychophragmus violaceus hybrids.

In an earlier study, the progenies of intergeneric hybrids Brassica napus (2n = 38) x Orychophragmus violaceus (2n = 24) were investigated in successive generations (F1-F4) for the cytological phenomenon of parental genome separation during mitotic and meiotic division. In the present study, inbred lines (F5-F8) derived from 1 such hybrid were characterized for morphology, chromosome pairing behaviour, and genome composition. One F5 plant (2n = 31) with slightly yellow petals and 12:19 and 15:16 segregation ratios in its pollen mother cells (PMCs) produced F6 plants with distinct morphological characteristics and wide variations in fertility and chromosome numbers (2n = 25-38). F7 and F8 lines with distinctive morphology and wide ranges in chromsome numbers were established. In PMCs of F7 plants from 4 F6 plants, 0-12 labelled chromosomes from O. violaceus, which predominantly appeared as bivalents, were identified by genomic in situ hybridization. They behaved synchronously with B. napus chromosomes during meiotic division. The results provide molecular cytogenetic evidence of the inclusion of O. violaceus chromosomes in the original hybrids and the cytology in the hybrids documented earlier. They also show that chromosome behaviour was altered and the parental chromosomes became synchronized after successive generations.