通过甘蓝型油菜和白芥属间杂种后代的小孢子培养获得二体异附加系

对3株甘蓝型油菜-白芥单体异附加系（2n=39）进行小孢子培养,共获得15株单倍体植株。经GISH鉴定,其中11株n=19,4株附加有一条白芥染色体（n=20）。经秋水仙素加倍处理,获得了二体异附加系(2n=40),这为远缘杂交后代外源基因的保存、纯合和利用提供了一条新的途径。     

芥白种间杂种后代多个性状的变异规律研究

通过种间杂交可以获得丰富的变异类型,创造许多新的种质资源。通过对芥菜型油菜和白菜型油菜种间杂交后代群体进行分析,研究杂种后代遗传变异规律。芥白杂种后代群体以偏白菜型和中间型植株为主,其余为偏芥菜型和其它新类型植株。随着自交世代的增加,花粉育性逐渐恢复,在BC, F3代已经趋于稳定。杂种后代变异广泛,还出现黄籽、自交亲和材料,且都是可遗传的。BC, F、群体叶色变异显著,SPAD值在10.6 }75.0之间变动,还出现叶色以及花器官突变体。芥白种间杂种后代群体变异丰富,拓宽了白菜型油菜的遗传基础。     

白菜型油菜黄籽沙逊和芥蓝种间杂交合成甘蓝型油菜研究

人工合成甘蓝型油菜是拓宽油菜遗传育种种质资源的有效途径,也是研究芸薹属物种起源与演化的重要手段之一。为了把黄籽沙逊的黄籽性状同芥蓝的优良特性相结合,本研究利用白菜型油菜黄籽沙逊(Brassica. campestris var. Yellow Sarson Pain, 2n=AA=20)与芥蓝(Brassica albograbra, 2n=CC=18)开展了芸薹属种间杂交研究,在MS+6-BA 1.0 mg/L+NAA 0.1 mg/L+7%蔗糖+活性炭1.0 mg/L的培养基上,通过胚珠培养从授粉7 d的140个胚珠中得到6个远缘杂交胚发育而成的植株,其中1个自然加倍成为双二倍体,随后应用形态学、细胞学和分子标记的方法对双二倍体种间杂种植株进行了鉴定,结果表明杂种植株生长势较强,形态属于中间类型;根尖染色体压片观察显示杂种植株具有父母本的染色体数量之和;SSR分子标记分析表明,杂种植株包含了双亲的遗传信息,由此证明了所获杂种的真实性,这为最后自交分离获得黄籽甘蓝型油菜提供了科学依据。     

甘蓝型黄籽油菜与黑籽油菜苗期生理特性的比较研究

油菜苗期的植株长势与抗性及后期产量密切相关。为探索甘蓝型黄籽油菜抗逆性较弱和产量较低的原因,以2对不同遗传来源的甘蓝型黄、黑籽油菜近等基因系为材料,研究了苗期植株的主要生理特性。结果表明,甘蓝型黄籽油菜苗期生长势弱,叶绿素含量和类胡萝卜素含量,光合速率和叶面积指数（LAI）均比相同遗传背景下的黑籽油菜低;与黑籽油菜相比,甘蓝型黄籽油菜无论是在不同的叶龄期,还是在植株的不同部位,均表现出“糖高氮低”的代谢特点,且越冬期叶片的硝态氮含量和硝酸还原酶活性也低;根颈粗和单株干物质重也明显小。甘蓝型黄籽油菜苗期生长势弱,冬前干物质积累偏少,进而影响其春后的生殖生长,这可能是其产量较低的重要原因之一。     

甘蓝型油菜雌性不育突变体FS1花器官形态结构研究

植物界中雌性不育现象普遍存在，迄今已在小麦、水稻、苎麻、油松、桔柑等作物中发现了雌性不育突变体^[1~5]，并对雌性不育的机理、遗传、应用进行了研究^[6~11]。1996年我们在油菜育种圃甘蓝型黄籽油菜宁油10号株系中发现了3株异常株，每株只结几个角果，角果肥大且角粒数只有1~5个，后来研究证实为雌性不育突变体，定名为FS1。雌性不育系作父本与母本混播，应用于杂交油菜制种可缩小父、母本间传粉距离，增加母本播种量，从而提高制种产量和纯度。初步研究表明，FS1天然授粉、套袋自交、剥蕾自交结角率为3%左右，平均每角粒数为1.30-2.07粒，其雌性不育性可以遗传，花粉育性正常，体细胞染色体数正常，为2n=38，花粉母细胞减数分裂正常（有关研究另文报道）。为了解甘蓝型油菜雌性不育突变体FS1的雌性不育机制，本文从形态结构方面做了初步研究。     

甘蓝型纯黄籽油菜新品系YR 5602的选育

通过甘白远缘杂交和复合杂交相结合的方法，经7年8代选育，于2004年育成了甘蓝型纯黄籽双低油菜新品系YR5602，其芥酸含量为1．18％～1．66％，硫甙28．8～38．3／μmol／g，含油量43．8％～45，2％，种子蛋白质23．3％～24．6％；群体黄籽株率达100％，黄籽频率达100％，黄籽度高达1．94～2．0，除种脐颜色稍深外，种皮全部为淡黄色且无斑点，不论是自交种或是天然种，其黄籽性状均表现极为稳定。已有两个甘蓝型油菜纯黄籽双低组合进入贵州省预备试验。至目前为止，象这样稳定的纯黄籽甘蓝型油菜双低高油分材料极为少见，是1份极为珍贵的重要种质资源，对甘蓝型油菜黄籽优质育种与杂种优势利用具有很高的利用价值．     

油菜及其近缘种种子萌发期耐盐碱性差异

为了明确甘蓝型油菜、白菜型油菜、芥菜型油菜、白芥和芸芥种子萌发期的耐盐碱性差异,分别采用0（CK）、0.4%、0.6%、0.8%、1.0%、1.2%和1.5%的NaCl溶液和0（CK）、0.1%、0.2%、0.3%、0.4%、0.5%和0.6%的NaHCO₃溶液进行萌发期胁迫处理,测定了发芽势、发芽率和发芽指数的变化规律。结果表明：不同浓度NaCl溶液处理,油菜及其近缘植物的平均发芽率依次为芥菜型油菜（71.7%）>甘蓝型油菜（64.0%）>白芥（62.4%）>白菜型油菜（48.4%）>芸芥（30.6%）。当NaCl的浓度为1.0%时,芥菜型油菜的发芽率最高,为91%,芸芥最低,仅有2%。不同浓度NaHCO₃溶液胁迫下,5种植物的平均发芽率依次为白芥（85.3%）>甘蓝型油菜（84.3%）>白菜型油菜（59.1%）>芸芥（42.9%）>芥菜型油菜（34.3%）。当NaHCO₃溶液浓度为0.4%时,白芥的发芽率最高,为95%,而芥菜型油菜最低,为0。综上,芥菜型油菜对NaCl胁迫的耐性最强,白菜型油菜和芸芥对NaCl胁迫的耐性差;白芥和甘蓝型油菜对NaHCO₃胁迫的耐性强,而芥菜型油菜对NaHCO₃胁迫的耐性较差。可见,在油菜种植区改良盐渍化土壤,中性盐渍化土壤最好选用芥菜型油菜,偏碱性盐渍化土壤可以选用白芥或甘蓝型油菜。     

甘蓝型黄籽油菜的遗传研究

从1975—1990年作者及其研究集体系统坚持了甘蓝型黄籽油菜的遗传育种研究。15年来取得了以下主要研究结果: 1.甘蓝型黄籽油菜的种皮色泽不同于白菜型、芥菜型和埃塞俄比亚油菜,为土黄或姜黄,而没有纯黄,只有杂黄,即在黄色种皮上有黑色斑点、斑块或褐色环带。 2.长期自交后仍得不到遗传上稳定的纯黄后代;在大群体中,不论是自     

甘蓝型黄籽油菜主要性状的通径分析

以7个不同遗传来源的甘蓝型黄籽品系配制的完全双列杂交组合作为样本，对甘蓝型黄籽油菜主要性状进行通径分析。主要结果为：全株有效果数和株高对单株粒重有最大正向作用，通径系数分别为0.514和0.535。皮壳率对黄籽度有直接的最大负效应，通径系数为-0.798。     

甘蓝型油菜与播娘蒿原生质体融合杂种后代的遗传研究

甘蓝型油菜(2n=38)与播娘蒿(2n=28)原生质体融合杂种F₁连续自交3代,获得F₂、F₃和F₄后代。用细胞学和SSR分子标记方法,分析杂种后代的染色体数目变异、减数分裂行为以及播娘蒿遗传成分的保留情况。结果表明在F₂、F₃和F₄代中,根尖细胞染色体平均数分别为38.47±3.17、37.65±3.23和36.66±2.95,随着自交世代增加呈减少趋势;在杂种后代减数分裂中,观察到染色体桥、染色体落后、染色体周期不同步、不均等分离等现象;杂种后代F₂、F₃和F₄代中检测到播娘蒿特征条带的平均频率分别为9.62%、2.99%和0.31%,呈减少趋势。因此要实现播娘蒿种质向油菜渗入应该重视F₂世代的选择。     

Cytogenetical and molecular investigations on somatic hybrids of Sinapis alba and Brassica juncea and their backcross progeny

Somatic hybrids of Sinapis alba+Brassica juncea (Sal Sal AABB) were synthesized by protoplast electrofusion. They were true genomic allopolyploids since they possessed 60 chromosomes, i.e. the sum of S. alba (2n= 24) and B. juncea (2n= 36) chromosomes. Chromosome pairing was predominantly bivalent with the occasional occurrence of multivalents in the pollen mother cells at diakinesis and metaphase I. Hybrids were completely pollen-sterile, but produced seeds on back-crossing with B. juncea and B. campestris. A total of 37 BC₁ plants were raised from two somatic hybrids (JS-1 and JS-2) and 24 of these were analysed cytologically. The 22 plants originating from the pollinations of somatic hybrids with B. juncea showed a chromosome configuration of 18II+12I and had 42–86% pollen fertility. Two plants from the backcrosses of the somatic hybrid with B. campestris formed 10II +20I, and had 0–4% fertile pollen. Total DNA analysis by probing with pTA71 carrying a full-length 18S–25S rDNA fragment of the wheat nuclear genome revealed that the two somatic hybrids possessed all the characteristic bands of both the species, confirming their hybridity. Probing with the mitochondrial coxI and atp9 genes indicated mitochondrial genome recombination in the hybrids. Hybridization with chloroplast-specific psbD indicated that both the somatic hybrids possessed the cp genome of S. alba origin.     

Hybridization of Sinapis alba L. and Brassica napus L. via Embryo Rescue

Embryo rescue techniques were used to obtain hybrids between Sinapis alba L. (white mustard) and Brassica napus L. (oilseed rape) with the goal of improving the disease tolerance of oilseed rape. Hybrid plants with 31 or 43 chromosomes were only recovered, when S. alba, was used as the female parent. One hybrid was obtained from the cross S. alba L. cv. ‘Kirby’×B. napus L. cv. ‘Topas’, while 26 hybrids were obtained, when various S. alba L. cultivars were pollinated with the rapid cycling B. napus line CrGC 5006. All F₁, hybrid plants were male sterile; however, the first generation backcross to B. napus L., also obtained by embryo rescue, produced plants with 50 chromosomes and 61–84 % pollen viability. Second backcross generation seed was produced by normal sexual crossing. Preliminary cytological analyses of pollen mother cells of hybrid plants suggests the possibility of genetic exchange between the two species.     

利用非条件和条件QTL解析油菜产量相关性状的遗传关系

基于前期研究中构建的Sollux/Gaoyou DH群体在9个环境中的表型数据和新版遗传图谱,对油菜角果长度进行QTL定位,估测QTL的加性、上位性和环境互作效应。并通过条件QTL方法,解析角果长度与角果粒数和粒重之间的遗传关系,以期利用标记辅助,探讨通过选择角果长度基因型以增加角果粒数、提高千粒重,最终达到增加产量的可能性。结果共检测到在3个环境以上稳定表达的控制角果长度QTL 8个,加性效应值在0.09~0.26 cm之间,效应总和解释群体遗传总变异的60%。8对上位性QTL效应值在0.035~0.075 cm之间,效应总和为加性总效应的38%。QTL与环境互作效应只在少数位点和个别环境中显著。条件QTL研究表明,q SLA2、q SLC1-2和q SLC8-1位点,角果长度的变化对角果粒数影响较大;而通过选择q SLA7、q SLC1-2、q SLC8-1和q SLC8-2长角果标记基因型,可望同时提高角果粒数和千粒重。6个主效QTL 11个连锁标记基因型和表现型的关联分析,验证了条件QTL分析结果,表明通过对q SLA2、q SLA7、q SLC8-1和q SLC8-2位点6个连锁标记(ZAAS423、SUC1-3、ZAAS12a、ZAASA7-28、ZAAS433和ZAAS437)长角果基因型的聚合,可增长角果约2 cm,间接增加角果粒数2粒,同时提高千粒重0.4 g,从而可望实质性地提高油菜产量水平。     

Genetic control of fatty acid composition in seed oil of Sinapis alba L.

Summary Inheritance of fatty acid composition was studied in an F1 diallel cross in Sinapis alba. Crosses were made among accessions having contrasting amounts of oleic (C18:1) and erucic (C22:1) acid. Concentrations of oleic, linoleic (C18:2), eicosenoic (C20:1) and erucic (C22:1) acids were determined by gas-chromatography for each mating combination. Genetic analysis confirmed that the composition of the fatty acids was controlled mainly by the nuclear genes of the embryo. Additive gene action with partial dominance for the reducing alleles was noted for oleic and linoleic acids, while erucic acid showed an additive mode of inheritance with partial dominance for the enhancing alleles. Positive heterosis was demonstrated for eicosenoic acid content. Erucic acid content was strongly negatively correlated with oleic acid, suggesting a genetic interdependence between the two fatty acids. Broad-sense and narrow-sense heritability estimates for each of oleic, linoleic and erucic acids were very high, due to low between-plants non-genetic component of variance.Contribution No. 3662-E, 1992 series.     

人工合成具有白菜或甘蓝细胞质的甘蓝型油菜

为比较甘蓝型油菜不同细胞质基因组的遗传效应及其与核基因的相互作用,对12个白菜(2n=20)与1个芥蓝(2n=18)的正反交种间杂种分别进行子房和胚培养,人工合成白菜和甘蓝细胞质甘蓝型油菜。结果表明,白菜×芥蓝的正交子房培养杂种苗平均诱导率2.32%,甘蓝×白菜的反交胚培养杂种苗平均诱导率为1.16%。不同杂交组合之间,杂种苗诱导率差异大,但相同亲本正反交杂种获得的难易趋势相似。将单倍体杂种小苗在含0.01%秋水仙碱的MS培养基中预培养处理10d的染色体加倍效果最好,加倍率达59.32%。人工合成的甘蓝型油菜农艺性状类似于栽培甘蓝型油菜,无论正交或反交合成的甘蓝型油菜,其农艺性状介于父母本之间,但更接近于母本。合成油菜花粉育性在40.53%～88.95%之间。     

甘蓝型油菜BnADH3基因的克隆及转BnADH3拟南芥的耐淹性

利用RT-PCR技术从甘蓝型油菜耐淹品系WR-4中克隆获得Bn ADH3基因,其完整开放阅读框为1137 bp。该基因编码379个氨基酸,与甘蓝Bo ADH3基因和拟南芥At ADH3基因高度同源,同源性分别达到96%和91%。利用定量RT-PCR检测Bn ADH3基因在油菜耐淹系WR-4和不耐淹系WR-24中的表达表明,在淹水处理下该基因表达受到一定的诱导,淹水处理6 h后表达开始上调,说明Bn ADH3基因在油菜耐淹机制中发挥作用;将其转化到模式生物拟南芥中,采用幼苗淹水3 d后去水处理,测定表明转基因株系叶片和根系中乙醇脱氢酶活性均高于野生型;生长4周和6周的拟南芥植株淹水3 d后的表型显示,Bn ADH3的表达可增强拟南芥对淹水胁迫的耐性,处理的T2代转基因幼苗大部分恢复,但野生型幼苗枯死;调查表明,淹水5 d后野生型植株的存活率为26.7%,转基因株系ADH33和ADH44的存活率分别为80.0%和66.7%。     

An evaluation of the potential of intergeneric gene transfer between Brassica napus and Sinapis arvensis

The possibility of gene transfer between Brassica napus and Sinapis arvensis was evaluated. Six spring-type cultivars of B. napus and four strains of S. arvensis were reciprocally crossed through controlled crosses. No hybrid was yielded from any cross. However, one hybrid with 28 chromosomes was obtained from B. napus×S. arvensis through ovule culture. The hybrid plant was highly sterile and set no seed on open pollination. Two F₂ plants, with 35 and 36 chromosomes respectively, were obtained through self-pollination by hand. Backcross of B. napus produced 23 plants carrying some characteristics of S. arvensis, but backcross to S. arvensis failed to produce a plant. The chromosome counts of the BC₁F₁ plants indicated that gametes with more than nine chromosomes were favoured during the meiosis. The data demonstrated that gene transfer from S. arvensis to B. napus was very difficult under controlled cross and backcross, while to transfer genes from B. napus to S. arvensis would be extremely remote even under the most favorable conditions.     

甘蓝型油菜株高性状的全基因组关联分析

适当的株高不仅是抗倒性和机械化收获的需要,而且是建立高产理想株型的需要。本研究测定来自世界油菜主产国的371份种质资源材料构成的自然群体的株高,利用60K SNP芯片对该群体进行基因型检测,获得了21 856个SNP,利用这些SNP和株高数据进行全基因组关联分析,结果表明:(1)检测到4个与株高显著关联的SNP,即Bn-A07-p3583161、Bn-scaff_22790_1-p1271170、Bn-scaff_20735_1-p42779和Bn-scaff_18702_1-p589589,分别分布于A07、C01和C02染色体;它们对表型变异的解释率分别为11.33%、11.75%、12.31%和10.97%;(2)参考基因组信息,在上述前3个SNP附近的492.0、9.5和69.5 kb处分别找到一个与株高性状相关的候选基因;(3)4个显著SNP各自的等位基因间表型均值差异分别为11.09、15.12、10.48和8.93 cm,双尾t测验结果表明各等位基因组间差异显著。