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%。     

甘蓝型油菜和芥菜型油菜种间杂交研究

甘蓝型油菜与芥菜型油菜杂交研究结果表明,杂交结实力与杂交组合方式以及参与杂交的亲本材料有关,以芥菜型油菜作母本的杂交结实力高于以甘蓝型作母本的组合：芥×甘杂交组合的平均结实数／花为 ２．６４ 粒,而甘×芥杂交组合的平均结实数／花为 ０．１０ 粒。芥甘杂种一代形态特征和生育期介于双亲之间,甘芥杂种一代不表现整齐的中间类型,株间差异明显;总体来看,芥甘杂种一代与双亲回交的结实力（０．４０,０．２１）低于甘芥杂种一代与双亲回交的结实力（３．３０,１．７４）,无论是芥甘杂种一代还是甘芥杂种一代,用甘蓝型油菜作父本回交的结实力高于用芥菜型油菜作父本回交的结实力,但也有个别回交组合出现例外,不表现上述规律。 Ｂ Ｃ１ 代种子当年播种出苗率低（１８．５％ ）,群体株间性状差异明显,生育期极不一致。芥甘杂种一代与甘蓝型油菜亲本第二次回交,其平均结实数／花较回交一代提高 １．０８ 粒, Ｂ Ｃ２ 代种子当年播种出苗率仍较低,但较对应的 Ｂ Ｃ１ 代稍有提高,群体中出现趋回交父本性状但雄性育性彻底退化的植株。芥甘杂种一代自由授粉所得 Ｆ２ 群体是一个变异极为丰富的遗传群体。     

芥菜型和甘蓝型油菜及其杂种后代种子硫甙组成差异的研究

用高效液相色谱技术分析了芥菜型油菜（Ｂｒａｓｓｉｃａ ｊｕｎｃｅａ）和甘蓝型油菜（Ｂｒａｓｓｉｃａ ｎａｐｕｓ）种间杂种后代及共亲本的硫甙组成。结果表明,与亲本农艺性状相似的后代,那么其硫甙组成与亲本亦相似。各后代中硫甙组成与亲本相比,虽发生了不同管理的变化,但都有倾向母体的趋势。芥菜型油菜烯丙基硫甙含量,２－羟基－３－丁烯基硫甙含量低,而甘蓝型油菜２－羟基３－丁烯基硫甙含量高,烯丙基硫甙含量最低     

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…     

芥菜型油菜和芥蓝种间杂种的获得及其性状表现

种间杂交是拓宽遗传种质资源,创制新物种、新材料的重要手段。利用芥菜型油菜和芥蓝种间杂交人工创制其种间杂种,并对其形态学、细胞学特点进行分析。结果表明:不同的杂交组合30 d后的子房残留数和获得幼胚数差异明显,杂交组合是影响种间杂交结子(幼胚)的重要因素;通过形态学鉴定出真杂种36份,均表现为较强的营养体杂种优势。真杂种减数分裂均为异常,在减数分裂后期具有不同程度的染色体丢失现象;无性系染色体数目部分为27条,部分为27~34条。真杂种花粉育性可染率在0~47.87%,自交结实率介于0~3.65%之间。     

白菜型油菜与蓝花子杂交的初步研究

通过胚胎培养,成功地获得了白菜型油菜（Ｂｒａｓｉｃａｃａｍｐｅｓｔｒｉｓ）与蓝花子（ＲａｐｈａｎｕｓｓａｔｉｖｕｓＬｖａｒｒａｐｈａｎｉｓｔｒｏｉｄｅｓＭａｋｉｎｏ）的属间杂种。该杂种具有两种类型;一种为大花类型,一种为小花类型。对它们进行花粉母细胞减数分裂的观察结果表明：小花类型为未加倍的杂种ＭＩ,存在１９个未配对染色体,大花类型为加倍或部分加倍杂种,加倍类型ＭＩ,１９个二价体排列在赤道板上;部分加倍类型ＡＩ,具有１０－１０－９的染色体组分割现象。大花类型具有可育性;它能够产生很多ｎ＝１９及ｎ＝９、ｎ＝１０的正常配子。染色体组分割能够产生倍半二倍体,它能用来研究染色体的功能和开展染色体工程。     

甘蓝型油菜和白菜型油菜种间杂种的小孢子培养

利用分离小孢子培养从甘蓝型油菜（Ｂｒａｓｓｉｃａ ｎａｐｕｓ）和白菜型油菜（Ｂ． ｃａｍ ｐｅｓｔｒｉｓ）种间杂种中获得了胚和再生植株。所用的培养程序是,将甘蓝型油菜和白菜型油菜杂种小孢子在蔗糖浓度为１７％ 、ＢＡ 为０．１ ｍ ｇ／Ｌ的液体ＮＬＮ 培养基中３２ ℃下暗培养４８ ｈ,再转入蔗糖浓度为１０％ 的ＮＬＮ 培养液中２５ ℃下暗培养３ 周。不同杂种间小孢子胚胎发生能力存在差异,其中ＵＭ９２１（白菜型油菜）×９１１１８６（甘蓝型油菜）正反交杂种的胚产量显著高于供试的其它组合。供体植株种植在１０ ℃／５ ℃（昼／夜）条件下能显著改善杂种小孢子胚产量和质量。杂种小孢子胚产量和杂种植株每荚种子数存在极显著正相关,但杂种植株的花粉育性和胚产量间相关不显著。大多数甘蓝型油菜和白菜型油菜杂种小孢子胚衍生植株为非整倍体,２２．８％ 的植株起源于具亲本染色体数的小孢子,几乎全部为ｎ＝ １９ 的类型。讨论了影响种间杂种小孢子胚胎发生的因素以及种间杂种小孢子培养技术的可能用途     

我国芥菜型油菜品种遗传多样性初探

用ＲＡＰＤ技术对包括春、冬芥菜型采及国外品种在内的３６个芥菜型油菜品种的遗传多样性进行了分析。在扩增得到的１２８条ＤＮＡ带中,多态性ＤＮＡ片段达８８．２８％。分析表明：春 油菜间遗传差异较大,国内冬性芥菜型油菜地方种多样性水平较高,２５份冬性品种分属Ⅰ、Ⅱ和Ⅲ类群,而春性芥菜型 采地方种均归于Ⅳ类;印度的ＲＬＭ１９８与四川的珙县金黄油菜、澳大利亚品种与我国春油菜品种亲缘关系密切。     

芥菜型多室油菜与甘蓝型油菜的种间远缘杂交

通过对芥菜型多室油菜与甘蓝型油菜种间杂交 以下简写为芥×甘或甘×芥 的结实性、交配性以及不同甘蓝型油菜对交配性的影响等研究发现:芥、甘正反交形成的饱满种子数较少,其形成种子的能力弱,但是芥×甘与甘×芥杂交相比,芥×甘形成饱满种子的能力较强,受精能力以及杂种胚胎的发育能力也强,在授粉后的子房发育上二者无显著差异.所以,芥菜型多室油菜与甘蓝型油菜种间杂交创建新资源时宜采用芥×甘杂交方式;不同甘蓝型油菜品种与芥菜型多室油菜正反交的结角率、受精指数、结籽指数和可交配指数均不相同,但可交配指数的变异系数最大.因此,筛选可交配性强的甘蓝型基因型应着眼于可交配指数高的甘蓝型油菜亲本材料,根据本试验结果,芥菜型多室油菜与甘蓝型油菜93-221-1杂交形成的杂种胚具有较强的可发育性.     

芥菜型油菜与埃塞俄比亚芥杂种基因组原位杂交分析

原产于非洲的埃塞俄比亚芥(Brassica carinata,2n=34,BBCC)具有适应于炎热干旱地区种植等特点,是改良我国芥菜型油菜(B.juncea,2n=36,AABB)的重要种质资源。本研究用基因组原位杂交方法(GISH,Genomic in situ hybridization)分析了芥菜型油菜与埃塞俄比亚芥种间杂种花粉母细胞的染色体分离,发现在后期I染色体主要以17∶18类型分离,其次是16∶19,染色体落后现象偶然发生,其中B染色体组以8∶8的分离比率较高,表明不同来源的B染色体可正常配对分离。本研究表明,芥菜型油菜与埃塞俄比亚芥远缘杂交,通过染色体同源重组(B染色体间),以及部分同源染色体配对交换的方式(A、B、C基因组间),有可能将埃塞俄比亚芥优良遗传成分转移到芥菜型油菜中。     

Phytoremediation of aspirin and tetracycline by Brassica juncea

With the increasing release of pharmaceutical drugs in the environment, research is in progress for investigating alternative methods for their remediation. Various studies have shown the phytoremediation potential of Brassica juncea for metals. The current study was aimed at evaluating the phytoremediation potential of B. juncea for two different pharmaceutical drugs i.e. aspirin and tetracycline in in-vitro conditions. The seeds of B. juncea were germinated and grown for a period of 28 and 24 days for aspirin and tetracycline, respectively. The study analyzed the remediation rate of B. juncea for the selected drugs in three different sets of varying concentration along with any phytotoxic effects exerted by the drugs on the seeds. Preliminary results showed that the average remediation rate of aspirin and tetracycline at the end of experiment was approximately 90% and 71%, respectively. As initial drug concentrations were increased in the media, the remediation rate also improved. However, at higher concentrations, the plants showed phytotoxicity as depicted by the decrease in shoot length of the germinated seeds. These preliminary results indicated that B. juncea could tolerate and remediate pharmaceutical drugs such as analgesics and antibiotics.     

Development of Plantlets from Leaf Protoplast Culture in Brassica juncea var. tsatsi

Protoplast of two mustard cultivars: Brassica juncea var. tsatsi cv. “Quxian Jiaoercai” and “Bangbangcai”, were isolated by enzymolysis from leaf grown in vitro. Protoplasts were suspended in liquid medium and semi-solidified medium with 0.35% low melting point agarose which formed a thin layer floating on the surface of the liquid medium. The first division appeared after 48h in the culture. One week after the original culture, a diluted medium with gradual dicrease of mannitol concentrations (6%→4%→zero) was then added to the culture three times respectively at one week's interval. In this culture method cell division and formation of microcalli were achieved. During the liquid culture of protoplasts, shaking at 20 rpm from time to time was beneficial in the formation of cell colonies and microcalli. Cell colonies developed into calli of approx 0.5—1mm in diameter one month after culture. The plating efficiency, which defined as the percentage of microcatli to numbers of protoplasts, was 0.2%—1%. Shoot regeneration occured when leaf protoplast-derived calli of “Quxian Jiaoercai” were transferred onto the modified MS medium supplemented with BAP 2.0mg/L, KT 1.0mg/L and NAA 0.2mg/L, and those of -'Bangbangcai" were transferred onto the modified MS medium supplemented with BAP 2.0mg/L. Individual shoot was rooted on a rooting medium supplemented with NAA 0.2 or 0.4 mg/L.     

Callus Formation and Morphogenesis of Cultured Mustard (Brassica juncea) Protoplasts

Protoplasts from mustard seedling cotyledons were suspended in Nitsch medium and cultured at 26℃ under low intensity illumination. When colonies were observed in the cultures, fresh medium (3% sucrose replaced mannitol) was added by equalvolume. When protoplasts developed into small callus the cultures were transfered to modified MS agar medium, in which some of them produced roots while an others gave rise to green spots. Shoot-differentiation experiment is in progress.     

大白菜细胞核复等位雄性不育基因定位

以大白菜不育材料"939A"为母本,携带恢复基因的材料YDQ56A为父本构建隐性雄性不育F1S4分离群体,利用混合分组分析法(BSA)对不育基因初定位;同时以"939A"为母本,携带保持基因的材料yellow sason为父本构建显性雄性不育F2分离群体,对不育基因进行精细定位。结果在F1S4群体中获得与不育基因连锁的标记2个,A08_1900和Br ID111035,与不育基因分别相距8.8c M和2.5c M,物理距离为804.1kb;F2群体中不育单株与可育单株分离比符合3∶1,不育基因表现为显性。通过标记验证,F2和F1S4两个群体定位结果一致,均位于A08染色体末端,通过新标记的筛选获得不育基因两侧紧密连锁的标记Br19470306和Br19675586,与不育基因分别相距1.6c M和2.4c M,物理距离205.28kb,其中包含58个基因。该结果为大白菜雄性不育系的利用以及转育奠定了基础。     

A RFLP-based linkage map of mustard [Brassica juncea (L.) Czern. and Coss.]

 A genetic linkage map of Brassica juncea was constructed based on restriction fragment length polymorphism (RFLP) detected by anonymous cDNA markers from B. napus, using a segregating F₁-derived doubled haploid (DH) progeny from a cross between a canola-quality mustard line (J90-4317) and a high-oil-content mustard line (J90-2733). The RFLP probes consisted of 229 cDNA probes from B. napus and a B. napus tandem repeat sequence, RDA2. The map consisted of 343 marker loci arranged in 18 major linkage groups plus five small segments with two to five marker loci, covering a total map distance of 2073 cM. Twenty-four percent of the markers were dominant in nature. Sixty-two percent of the marker loci were duplicated, and the majority were involved in inter-linkage group duplications, illustrating that complex duplications and subsequent rearrangements occurred after allopolyploidy. Deviation from the Mendelian segregation ratio for a DH population was observed for 27% of the markers. Two-thirds of these markers with a skewed segregation were clustered in 6 linkage groups and two unassigned segments. The overall average marker interval of the B. juncea map reported here was 6.6 cM, which would provide a marker density satisfactory for efficient use of the map in breeding applications, such as tagging of important agronomic traits and marker-assisted selection. Received: 14 May 1996 / Accepted: 11 October 1996     

Involvement of Asada-Halliwell Pathway During Phytoremediation of Chromium (VI) in Brassica juncea L. Plants

Brassica juncea (Indian mustard) L. plants were exposed to different concentrations (0.0, 0.1, 0.3 and 0.5 mM) of Chromium (Cr) and harvested after 30 and 60 days of sowing for the analysis of growth parameters, metal uptake and oxidative stress markers. Significant accumulation of Cr (VI) by B. juncea L. plants resulted in the reduced growth and modulations in the pool of various biochemical stress markers. The toxic effects of Cr (VI) on growth and other stress markers (protein content, lipid peroxidation and antioxidative enzymes viz.SOD, CAT, POD, APOX, GR, DHAR and MDHAR) in B. juncea L. were observed to be concentration and time dependent. Effect of Cr (VI) on biochemical parameters was differential and their maximum activities of SOD, POD, APX, GR, DHAR and lipid peroxidation were recorded at 0.5 mM concentration in 30 days old plants. Whereas, trend in the activities of most of the stress markers was reversed in 60 days old plants. The results obtained from the study suggested that Cr (VI) stress inhibited growth of B. juncea L. plants is directly interrelated with its accumulation and resulted in the modulation in activities of various stress markers.