Intergeneric Somatic Hybridization Between Brassica napus L. and Sinapis alba L.

Electrically induced protoplast fusion was used to produce somatic hybrids between Brassica napus L. and Sinapis alba L. Seven hybrids were obtained and verified by the simple sequence repeat and cleaved amplified polymorphic sequence analysis of the genefael, indicating that the characteristic bands from S. alba were present in the hybrids. The hybridity was also confirmed by chromosome number counting because the hybrids possessed 62 chromosomes, corresponding to the sum of fusion-parent chromosomes. Chromosome pairing at meiosis was predominantly normal, which led to high pollen fertility,ranging from 66% to 77%. All hybrids were grown to full maturity and could be fertilized and set seed after self-pollination or back-crosses with B. napus. The morphology of the hybrids resembled characteristics from both parental species. An analysis of the fatty acid composition in the seeds of F1 plants was conducted and the seeds were found to contain different amounts of erucic acid, ranging from 11.0% to 52.1%.     

甘蓝型油菜与紫罗兰属间杂交的植物遗传学研究

为探索属间杂种的遗传特点以及改良甘蓝型油菜油分品质,进行了甘蓝型油菜和紫罗兰的属间杂交.杂交母本为甘蓝型油菜奥罗(Brassica napus L. cv. oro),父本为紫罗兰(Matthiola incana (L.) R. Br.).将授粉7 d后的油菜子房切下,消毒后,培养于添加适当植物激素的MS培养基.从750个离体培养的油菜授粉子房中,获得了2粒成熟胚胎,其结籽率为0.26%.将胚胎取出,转接于MS培养基(添加2.0 mg/L 6-BA和0.1 mg/L NAA),获得了丛生芽.将丛生芽分割为许多单芽,转接到新鲜培养基中,长成了22株小苗.杂种一代植株呈中间性,它的许多性状介于两个亲本之间,一些性状倾向母本,少数性状表现显著的超亲杂种优势,植株结实性差.杂种后代(F2)植株表现多样性,多数植株的性状倾向母本,能育.部分植株表现中间性、育性差,少数植株发育不良、不育.染色体研究表明,杂种一代植株为混倍体.在杂种体细胞中,许多细胞的染色体数为2n=26,为两个亲本的配子染色体数之和.杂种后代(F2)中,倾母植株的染色体数为2n=38,矮小植株的许多细胞具非整倍染色体数,如2n-1=37、2n+1=39.从杂种后代中获得了种子油分品质较好的植株,有可能用于油菜的品质育种.     

甘蓝型油菜与紫罗兰属间杂交的植物遗传学研究

为探索属间杂种的遗传特点以及改良甘蓝型油菜油分品质,进行了甘蓝型油菜和紫罗兰的属间杂交。杂交母本为甘蓝型油菜奥罗（Brassica napusL.cv.oro)。父本为紫罗兰（Motthiola incana(L.)R.Br.)。将授粉7d后的油菜子房切下,消毒后,培养了添加适当植物激素的MS培养基。从750个离体培养的油菜授粉子房中,获得了2粒成熟胚胎,其结籽率为0.26%。将胚胎取出,转接于MS培养基（添加2.0mg/L6-BA和0.1mg/LNAA）,获得了丛生芽,将丛生芽分割为许多单芽,转接到新鲜培养基中,长成了22株小苗,杂种一代植株呈中间性,它的许多性状介于两个亲本之间,一些性状倾向母本,少数性状表现显著的超亲杂种优势,植株结实性差。杂种后代（F2）植株表现多样性,多数植株的性状倾向母本,能育。部分植株表现中间性,育性差,少数植株发育不良,不育,染色体研究表明,杂种一代植株为混倍体。在杂种体细胞中,许多细胞的染色体数为2n=26。为两个亲本的配子染色体数之和。杂种后代（F2）中,倾母植株的染色体数为2n=38。矮小植株的许多细胞具非整倍染色体数。如2n-1=37,2n 1=39。从杂种后代中获得了种子油分品质较好的植株,有可能用于油菜的品质育种。     

甘蓝型油菜芥酸含量的主基因＋多基因遗传

应用植物数量性状主基因＋多基因混合遗传模型对甘蓝型油菜无芥酸品种HSTC     

人工合成甘蓝型油菜中花色与芥酸含量的遗传连锁分析

利用人工合成的甘蓝型油菜品系．No．2127-17(白花、有芥酸)与加拿大双低甘蓝型油菜品种Quantum(黄花、低芥酸)配制杂交组合。对亲本、F1、BC1、F2和DH(doubled haploid)5个世代的花色及芥酸含量进行分析,结果表明：花色受单基因控制,且白花对黄花为显性;芥酸含量仅表现出一对基因的差异且具有加性效应的遗传模式。花色和芥酸含量的连锁分析表明：白花与高芥酸紧密连锁．在DH群体中重组频率为5．8％。采用集团分离分析法(Bulked Segregant Analysis,BSA),从685条10个碱基的随机引物筛选到一个与黄花和低芥酸含量紧密连锁的RAPD标记S92-1400。在遗传图谱上黄花基因和低芥酸基因距离S92-1400标记的图距分别为2．2cM和5．4cM。     

Detection of 5S and 25S rRNA genes in Sinapis alba, Raphanus sativus and Brassica napus by double fluorescence in situ hybridization

Different ribosomal RNA (5S and 25S) genes were investigated simultaneously by fluorescence in situ hybridization (FISH) in Sinapis alba, Raphanus sativus and Brassica napus. The chromosomes of S. alba carried four 5S and six 25S gene sites, and those of R. sativus four sites of each gene, respectively. These two species have one chromosome pair with both rDNA genes; the two are closely located on a short arm of S. alba, while in R. sativus one is distal on the short arm (5S) and the other more proximal on the long arm (25S). In B. napus we have confirmed 12sites of 25S rDNA. The detection of 5S rDNA genes revealed 14 signals on 12 chromosomes. Of these, six chromosomes had signals for both rDNA genes. The FISH with 5S rDNA probes detected two sites closely adjacent in four chromosomes of B napus. These results are discussed in relation to a probable homoeologous chromosome pair in B. oleracea. Received: 20 July 1999 / Accepted: 8 October 1999     

甘蓝型油菜与芝麻菜体的细胞杂交

为拓宽油菜育种的基因资源库, 改良油菜品种, 以甘蓝型油菜(Brassica napus)花油3号下胚轴和芝麻菜(Eruca sativa)下胚轴为材料分离制备原生质体; 然后采用PEG-高Ca2+-高pH法进行原生质体融合, 当PEG浓度为35%, 原生质体融合密度为5×105个/mL时, 融合25 min时, 融合率可达18.2%。融合后在培养密度为1×105个/mL时, 以附加1.0 mg/L 2,4-D +0.5 mg/L 6-BA+0.5 mg/L NAA+ 200 mg/L肌醇+300 mg/L水解酪蛋白的改良的KM8p为融合体培养基, 以0.1 mol/L 蔗糖+0.2 mol/L葡萄糖+0.2 mol/L甘露醇作渗透稳定剂进行液体浅层培养, 效果较好, 愈伤组织再生率最高为6.8%。将融合体再生的小愈伤组织转移至培养基(B5无机盐+0.087 mol/L蔗糖+0.2 mg/L 2, 4-D+0.5 mg/L NAA+0.2 mg/L 6-BA+ 0.5% Agar, pH 5.8)上增殖培养, 待愈伤组织长至直径为3~5 mm时, 及时将其转至分化培养基(MS无机盐+0.087 mol/L 蔗糖+0.1 mg/L IAA+0.8 mg/L 6-BA+0.8% Agar, pH 5.8)中诱导不定芽再生, 芽分化率为35.7%。当不定芽长为2~3 cm时, 将其切下转入附加0.5 mg/L IBA+0.2 mg/L 6-BA的1/2MS生根培养基中诱导生根, 14 d左右即可形成再生植株, 生根率可达88%。同时, 以紫外线(60 μW/cm2)照射芝麻菜原生质体, 进行不对称融合, 照射2 min的获得了愈伤组织和再生植株, 照射4 min的只获得愈伤组织, 而照射5 min以上的没有获得愈伤组织, 但其愈伤组织再生、增殖及植株再生均不如对称融合。从细胞学鉴定的21块杂种愈伤组织上再生出16株杂种植株。     

甘蓝型油菜与芝麻菜体的细胞杂交

为拓宽油菜育种的基因资源库, 改良油菜品种, 以甘蓝型油菜(Brassica napus)花油3号下胚轴和芝麻菜(Eruca sativa)下胚轴为材料分离制备原生质体; 然后采用PEG-高Ca2+-高pH法进行原生质体融合, 当PEG浓度为35%, 原生质体融合密度为5×105个/mL时, 融合25 min时, 融合率可达18.2%。融合后在培养密度为1×105个/mL时, 以附加1.0 mg/L 2,4-D +0.5 mg/L 6-BA+0.5 mg/L NAA+ 200 mg/L肌醇+300 mg/L水解酪蛋白的改良的KM8p为融合体培养基, 以0.1 mol/L 蔗糖+0.2 mol/L葡萄糖+0.2 mol/L甘露醇作渗透稳定剂进行液体浅层培养, 效果较好, 愈伤组织再生率最高为6.8%。将融合体再生的小愈伤组织转移至培养基(B5无机盐+0.087 mol/L蔗糖+0.2 mg/L 2, 4-D+0.5 mg/L NAA+0.2 mg/L 6-BA+ 0.5% Agar, pH 5.8)上增殖培养, 待愈伤组织长至直径为3~5 mm时, 及时将其转至分化培养基(MS无机盐+0.087 mol/L 蔗糖+0.1 mg/L IAA+0.8 mg/L 6-BA+0.8% Agar, pH 5.8)中诱导不定芽再生, 芽分化率为35.7%。当不定芽长为2~3 cm时, 将其切下转入附加0.5 mg/L IBA+0.2 mg/L 6-BA的1/2MS生根培养基中诱导生根, 14 d左右即可形成再生植株, 生根率可达88%。同时, 以紫外线(60 μW/cm2)照射芝麻菜原生质体, 进行不对称融合, 照射2 min的获得了愈伤组织和再生植株, 照射4 min的只获得愈伤组织, 而照射5 min以上的没有获得愈伤组织, 但其愈伤组织再生、增殖及植株再生均不如对称融合。从细胞学鉴定的21块杂种愈伤组织上再生出16株杂种植株。     

甘蓝型油菜与蔊菜的原生质体融合与植株再生

以甘蓝型油菜下胚轴和蔊菜叶片为外植体提取原生质体, 采用PEG-高pH、高Ca²⁺附加DMSO的原生质体融合方法, 用液体浅层静置培养融合体, 获得了10株融合杂种, 观察了杂种形态学和细胞学。结果表明：1%纤维素酶+0.2%离析酶+3 mmol/L MES 酶解14 h 可获得较高产率的油菜原生质体, 0.25% 纤维素酶+0.5%离析酶+5 mmol/L MES酶解12 h可获得较高产率的蔊菜原生质体; 30% PEG + 0.3 mol/L葡萄糖+50 mmol/L CaCl₂•2H₂O +15%DMSO的融合条件下, 获得了10.4%的融合率; 实验所获的原生质体融合材料可作为新种质。     

甘蓝型油菜花粉离体培养

通过对甘蓝型油菜花粉发育阶段和活力的检测确定花粉发育的时期,分离出单核晚期花粉进行离体培养.结果表明,(1)筛选出适合油菜小孢子花粉离体培养的液体培养基为T_1+怀特维生素(White's vitamins)+2%椰子汁+0.5 mol/L麦芽糖,在此培养基上花粉的成熟率可达25.1%,萌发率达6.3%.(2)筛选出适合成熟花粉离体萌发液体培养基为0.6 mol/L麦芽糖+1.6 mmol/L硼酸+2.9 mmol/L硝酸钙+29.6 μmol/L VB_1,在此培养基上,自然成熟花粉的萌发率可达75.2%.将离体培养成熟的花粉培养在萌发培养基,萌发的花粉占成熟花粉的66.3%.     

Scanning electron microscopy of desiccation-tolerant and -sensitive microspore-derived embryos of Brassica napus L.

Desiccation tolerance can be induced in microspore-derived embryos of Brassica napus L. by application of abscisic acid (ABA). Scanning electron microscopy was employed to study and compare desiccation-tolerant and -sensitive microspore-derived embryos. The external surface of those desiccated embryos in which desiccation tolerance had been induced was uniformly shriveled due to severe dehydration, and their internal tissue system was well preserved. In contrast, in desiccation-sensitive embryos, dehydration caused tearing of the epidermis and collapse of the internal tissue system. After the desiccated embryos had been rehydrated, the size and external morphology of the desiccation-tolerant ones recovered to the pre-desiccation state within 1 day, whereas the sensitive ones did not recover or remained shriveled. The effect of ABA on the induction of desiccation tolerance is discussed. Received: 25 June 1998 / Revision received: 03 September 1998 / Accepted: 24 September 1998     

Interspecific somatic hybridization between Brassica napus and Brassica hirta (Sinapis alba L.)

Summary Somatic hybridization between Brassica napus and B. hirta (or Sinapis alba) is described. No cybrid plant with B. napus nucleus exhibiting cytoplasmic male sterility was recovered. Somatic hybrids were identified morphologically and, for some of them, by cytological observations. They were also characterised by Southern hybridization of nuclear rDNA. Chloroplast and mitochondrial DNA restriction analysis showed that 2 plants out of 14 have B. hirta ctDNA, one the B. napus mtDNA and the other a hybrid. Nine possess B. napus ctDNA with a hybrid mtDNA. For six of them, mtDNA patterns present novel bands, suggesting intergenomic recombination during fusion. These hybrids will be included in the breeding program.     

甘蓝型油菜小孢子培养技术的几项改进

本研究在NLN-16和NLN-13的培养基中分别加入0.1mg/L 6-BA和0.05%的活性碳,结果表明6-BA对小孢子再生胚有明显地促进作用,再生胚的频率比对照增加26枚/皿,经分析达到显著水平;而0.05%的活性碳对小孢子再生胚促进作用不显著。对甘蓝型油菜小孢子培养再生植株的染色体加倍及移栽的研究结果表明,在小孢子培养初期加50mg/L秋水仙碱加倍效率最佳,加倍率达到67.6%。小孢子培养的再生苗移栽至大田后,采用遮阳网覆盖小苗,移栽成活率达到87.6%。 Abstract:The application of microspore culture technique was restricted because of its low frequency of embryogenesis and chromosome doubling.Two methods of enhancing the frequency of embryogenesis were employed in the study,namely,activated charcoal treatment in NLN-13 media and 6-BA treatment in NLN-16 media.The treatment with 0.05% activated charcoal produced 24 embryos per plate,which increased 1.7 embryos per plate,as compared with the treatment without activated charcoal.However,the analysis of T-test showed that it was not significant.After adding 0.1mg/L 6-BA in NLN-16 media,the frequency of embryogeny was 38.3 embryos per plate,and it was 26 embryos more per plate than that of CK.Analysis of T-test is significant.This indicates that 6-BA promotes embryogeny in microspore culture.Adding 50mg/L colchicines in NLN-16 media,the doubling frequency was 67.6%.The plantlets transplanted into field with two methods of light-covered net and plastic films were investigated.A survival rate of 87.6% was obtained using light-covered method whereas 57.7% survived using plastic film method.     

转入HPT1基因的油菜种子中维生素E含量的提高

尿黑酸植基转移酶（HPT）是天然维生素E合成途径中的关键酶,拟南芥中编码HPT的基因HPT1转入油菜种子特异表达后得到转基因油菜。初步的检测发现,其种子中的维生素E含量有所提高。     

油菜雄性核不育系及其等位可育系小孢子发育过程的比较研究

用Olympus BH2型光学显微镜对甘蓝型油菜单显性核不育系(GMS)及其等位可育系小孢子发育过程进行解剖学观察,发现在正常小孢子发育过程中,绒毡层在小孢子发育的四分体前后开始解体,为小孢子继续发育提供营养,而不育系小孢子的败育在减数分裂前就已经发生,并且不能形成四分体,小孢子逐渐解体,且小孢子解体在绒毡层解体之前发生,最后花药成为干瘪的空壳,导致不育。     

甘蓝型油菜若干农艺性状与单株产量的关系分析

以38个甘蓝型油菜品种为材料,对单株产量、有效分枝部位高度、主轴花序长度等11个农艺性状进行了方差分析、通径分析和聚类分析．结果显示:所有调查性状在品种间存在显著的差异;主花序长度、主序有效角果数与单株产量都有显著的遗传相关性,两者都通过有效分枝部位高度对单株产量产生较大的正向间接作用;有效分枝部位高度对单株产量的影响也较显著,高产品种具有相对较高的有效分枝部位高度;总角果数对单株产量的直接遗传通径系数虽小,但它具有最大的直接表型通径系数,它通过主序有效角果数和有效分枝数对单株产量产生了两个较大的间接作用;要获得3130kg/hm~2的产量,重点应具备果多、枝多的特征,总角果数不少于657个,有效分枝数不少于18个．     

甘蓝型油菜温敏不育系417S育性遗传及RAPD分析

通过对新选育的甘蓝型油菜高温敏感生态型不育系417S与30份中外品种(系)进行测交、杂交和回交,研究其温敏不育性的遗传特点.结果表明:(1)30份品种(系)可不同程度的恢复417S的育性,其中24个品种(系)可完全恢复417S育性,占所配组合的80.0%;5个品种(系)可高度恢复417S的不育性,占所配组合的17.2%.(2)自然条件下同一组合正反交F2群体育性分离结果不同,人工控温条件下不同组合F2分离群体中可育与不育呈现15∶1分离,回交群体中出现3∶1分离,表明417S温敏不育性受细胞质和2对隐性重叠核基因共同控制.(3)以417S与1521C构成的回交一代群体为试材,采用分离群体分组分析法(BSA法)对回交一代群体构成的不育集团和可育集团筛选育性基因的RAPD标记,从247个供试引物中筛选出2个引物BA392(5′-AGTCACTCCC-3′)、S113(5′-GACGCCACAC-3′)在不育群体和可育群体间扩增出多态性产物.经进一步对F2分离群体的单株检测,引物BA392扩增出了得到重复验证和可靠的多态性扩增产物,其产生的特异条带BA392-400bp与417S温敏雄性不育的恢复基因连锁,遗传图距为6.0cM,可作为417S温敏不育恢复基因的连锁标记.     

Hypocotyl growth in Sinapis alba L: the roles of light quality and quantity

Abstract. A comparison is made of the relative effectiveness of light quality and light quantity on the elongation growth of Sinapis alba hypocotyls. The results show that hypocotyl extension rate in plants which have not previously been exposed to light is controlled primarily by the prevailing photon fluence rate when the phytochrome photostationary state lies between ∼0.033 and ∼0.81. Below ∼0.033, changes in photostationary state also have a marked effect on extension rate. Elongation growth in light-adapted plants is controlled by both photon fluence rate and the spectral quality of the incident radiation at all photoequilibria. Photosynthesis can modify these responses but is not essential as a prior condition for a green plant to respond to changes in light quality and quantity.     

白芥自交亲和性分析

对不同来源的8份白芥材料,采用人工自交法分析其自交亲和性。结果显示:白芥的自交亲和性存在较大幅度的变异,自交亲和指数在0.01~4.10之间,8份参试材料中,自交亲和指数小于1的材料有3个,自交亲和指数大于1的材料有5个。表明白芥中存在自交亲和材料,白芥自交亲和性变异不仅存在于材料间,而且也存在于同一材料内不同个体间。按自交亲和指数的高低,可将参试材料分为3种类型:高自交亲和类型(自交亲和指数大于3.00,如民乐洪水芥麻、04(X)等)、自交亲和类型(自交亲和指数为1.00~2.99)、自交不亲和类型(自交亲和指数0.00~1.00)。     

Isozyme analysis as a tool for introgression of Sinapis alba germ plasm into Brassica napus

Summary Isozyme analysis of Brassica napus cv Topas and CGRC5006 as well as of Sinapis alba cv Emergo revealed significant polymorphism between the two species for the isozymes, aconitate hydratase, glucose phosphate isomerase, and diaphorase. F₁ hybrids between B. napus 5006 and S. alba cv Emergo were backcrossed to B. napus cv Topas, and the S₁ progeny of the first two backcrosses were studied isozymically. At the backcross one level the frequency of S. alba or S. alba plus B. napus patterns observed ranged from 18% to 87% across the four lines studied. There were differences between lines for the frequency of S. alba patterns, which could have an impact on the efficiency of selection for subsequent backcrossing. By the backcross two generation in one of the two lines studied, GR86-24, the S. alba patterns for GPI and DIA had been lost, while in the other line, GR86-28, the S. alba pattern for ACO had been lost, resulting in lost opportunity for S. alba gene transfer. In a wide cross such as S. alba x B. napus, which requires an intensive effort to accomplish, the isozymes ACO, GPI, and DIA may serve as useful markers to ensure gene transfer between the two species has occurred. In addition, the identification of lines with divergent isozyme patterns from B. napus will provide the basis for establishing linkages between S. alba traits of interest and isozyme markers.