菜薹-芥蓝单体异附加系n+1配子传递及两个二体异附加系的获得

【目的】研究菜薹-芥蓝单体异附加系n+1配子传递,为其在基因定位和遗传改良上的应用奠定基础。【方法】通过观察和统计各单体异附加系减数分裂后期Ⅱ10/10/11/11分离的PMCs,估算n+1配子的形成频率;通过鉴定回交子代植株的染色体数目,测定各单体异附加系n+1配子的传递率;从单体异附加系的自交子代中筛选二体异附加系。【结果】依据减数分裂后期Ⅱ10/10/11/11分离的PMCs比率估算,各单体异附加系n+1雄配子的形成频率在36.85%—45.15%;基于染色体数目鉴定,各单体异附加系n+1雄配子的传递率在7.14%—14.81%,雌配子在19.70%—36.51%;在一定范围内,n+1配子传递率与n+1配子形成频率、染色体大小、花粉量和花粉生活力没有显著的相关性;从自交子代中获得了两个不同的二体异附加系。【结论】菜薹-芥蓝各单体异附加系的额外染色体通过雌、雄配子均能进行传递,但传递率因不同的附加系而异;从单体异附加系的自交子代中可以分离出遗传性稳定的二体异附加系。     

Monosomic Addition Lines of Flowering Chinese Cabbage （B. campestris L. ssp. chinensis var. parachinensis L. H. Bailey）-Chinese Kale （B. oleracea L. var. alboglabra L. H. Bailey）

Interspecific alien addition lines have played significant roles in gene mapping, intergenomic gene transfer and chromosomal homoeological identification between closely related species. Selection of alien addition lines was conducted by karyotype analysis and morphological observation with the reference of parents. Triploid interspecies hybrid （AAC, 2n = 3x = 29） was obtained from Brassica campestris ssp. chinensis var. parachinensis Qinglu 9601 （tetraploid, AAAA, 2n = 4x = 40） x B. oleracea var. alboglabra Baihua 9705 （diploid, CC, 2n = 2x = 18） by immature hybrid embryo culture in vitro. Five different alien monosomic addition lines （AA ＋ C2, AA ＋ C3, AA ＋ C4, AA ＋ C6, AA ＋ C7） were obtained from the backcross progenies of AAC x AA. Each alien monosomic addition line has some specific morphological characters. It is feasible to obtain alien addition lines from the progenies of AAC × AA by karyotype analysis and morphological observation based on the reference of parents     

大白菜-结球甘蓝双单体异附加系的SSR鉴定及其特性

 【目的】筛选大白菜-结球甘蓝异附加系,利用SSR标记鉴定附加的外源染色体,并对异附加系特性进行研究。【方法】以大白菜-结球甘蓝异源三倍体（AAC）与二倍体大白菜（AA）回交一代的自交后代为试材,利用根尖染色体计数、甘蓝连锁群特异SSR标记等方法,筛选、鉴定大白菜-结球甘蓝异附加系。【结果】在BC1F2中筛选到2n=22的植株;该植株对位于甘蓝01连锁群的3对SSR引物和08连锁群的2对SSR引物的扩增结果与异源三倍体杂交种是一致的,而对位于其它7个连锁群的20对特异引物的扩增结果与大白菜一致,表明该植株附加的外源染色体与甘蓝的01、08连锁群相对应,为双单体异附加系;该双单体异附加系绝大多数花粉母细胞减数分裂终变期染色体构型为10Ⅱ+2Ⅰ,后期Ⅰ以11-11、10-12分离方式为主,后期Ⅱ产生了n=11配子。【结论】利用甘蓝连锁群相对于大白菜特异的SSR标记可以准确鉴定大白菜-结球甘蓝异附加系;获得了附加甘蓝01、08连锁群的大白菜双单体异附加系,为进一步合成附加甘蓝01或08连锁群大白菜-结球甘蓝单体和双体异附加系奠定基础。     

Monosomic Addition Lines of Flowering Chinese Cabbage (B. campestris L. ssp. chinensis var. parachinensis L. H. Bailey)-Chinese Kale (B. oleracea L. var. alboglabra L. H. Bailey)

Interspecific alien addition lines have played significant roles in gene mapping, intergenomic gene transfer and chromosomal homoeological identification between closely related species. Selection of alien addition lines was conducted by karyotype analysis and morphological observation with the reference of parents. Triploid interspecies hybrid (AAC, 2n=3x=29) was obtained from Brassica campestris ssp. chinensis var. parachinensis Qinglu 9601 (tetraploid, AAAA, 2n=4x=40)×B. oleracea vat. alboglabra Baihua 9705 (diploid, CC, 2n=2x=18) by immature hybrid embryo culture in vitro. Five different alien monosomic addition lines (AA C2, AA C3, AA C4, AA C6, AA C7) were obtained from the backcross progenies of AAC×AA. Each alien monosomic addition line has some specific morphological characters. It is feasible to obtain alien addition lines from the progenies of AAC×AA by karyotype analysis and morphological observation based on the reference of parents.     

大白菜-结球甘蓝5号二体异附加系的选育及鉴定

【目的】获得稳定遗传的大白菜-结球甘蓝二体异附加系。【方法】以大白菜-结球甘蓝5号单体异附加系为材料,对其自交后代进行细胞学及SSR鉴定。【结果】37株自交后代中2n=22植株的比率为10.81%,通过对4个2n=22的植株进行核型分析及SSR鉴定,确定其为大白菜-结球甘蓝5号二体异附加系。减数分裂观察发现,大白菜-结球甘蓝5号二体异附加系后期Ⅰ虽然出现染色体落后现象,但在后期Ⅱ中染色体的分裂方式以11-11-11-11为主,其比率为69.55%。【结论】获得了大白菜-结球甘蓝5号二体异附加系,其外源染色体与甘蓝07连锁群相对应;该二体异附加系减数分裂形成的四分体以附加1条的染色体为主(n=11),有较高的遗传稳定性。二体异附加系的获得为研究芸薹属A、C基因组的亲缘关系,以及向大白菜中导入结球甘蓝的优良基因具有重要意义。     

Assignment of unanchored scaffolds in genome of Brassica napus by RNA-seq analysis in a complete set of Brassica rapa-Brassica oleracea monosomic addition lines

The economically valuable oil crop Brassica napus(AACC, 2 n=38), which arose from interspecific hybridization between the diploid ancestors Brassica rapa(AA, 2 n=20) and Brassica oleracea(CC, 2 n=18), has a complex genome. More than 10% of the assembled sequences, most of which belong to the C subgenome, have not been anchored to the corresponding chromosome. Previously, a complete set of monosomic alien addition lines(MAALs, C1–C9) with each of the nine C-subgenome chromosomes added to the extracted A subgenome was obtained from the allotetraploid B. napus donor Oro, after the ancestral B. rapa(RBR Oro) genome was restored. These MAALs effectively reduced the complexity of the B. napus genome. Here, we determined the expression values of genes on unanchored scaffolds in the MAALs and RBR Oro. Then, multiple comparisons of these gene expression values were used to determine the affiliations of the nonanchored scaffolds on which the genes were located. In total, 54.68%(44.11 Mb) of the 80.67 Mb of non-anchored scaffolds belonging to the C subgenome were assigned to corresponding C chromosomes. This work highlights the potential value of these MAALs in improving the genome quality of B. napus.     

栽培稻与紧穗野生稻回交后代的细胞学与褐飞虱抗性鉴定

】对栽培稻０２４２８与原产于乌干达的紧穗野生稻的ＢＣ２Ｆ１和ＢＣ１Ｆ２世代植株进行了细胞学研究,分离出单体异源附加系１５５份和易位系１８４份;抗褐飞虱特性鉴定表明,２８个单体异源附加系和４８个易位系表现抗虫。     

利用PCR技术初步鉴定小麦-加州野大麦异染色体系

为快速鉴定普通小麦与普通小麦—加州野大麦双二倍体杂交、回交后代植株的染色体组成，研究小麦背景中添加的外源染色体与小麦染色体之间的部分同源关系，选用已被定位在小麦7个部分同源群21条染色体上的38个SSR引物对杂种回交后代植株进行PCR扩增。结果表明，其中27个小麦SSR引物在普通小麦与小麦—加州野大麦双二倍体间有多态性扩增，涉及4个部分同源群的11对引物，可在不同杂种回交植株中扩增出与双二倍体相同的多态带纹；根据PCR扩增和细胞遗传学分析的结果，在18个回交后代中初步鉴定出7个可能的异附加系，其中2个二体异附加系、1个端二体异附加系、2个单体异附加系和2个双单体异附加系。所选育的异附加系分别涉及第1、2、4和7部分同源群。     

A Co-Dominant Marker BoE332 Applied to Marker-Assisted Selection of Homozygous Male-Sterile Plants in Cabbage (Brassica oleracea var. capitata L.)

The dominant genic male sterility (DGMS) gene CDMs399-3 derived from a spontaneous mutation in the line 79-399-3 of spring cabbage (Brassica oleracea var. capitata L.), has been successfully applied in hybrid seed production of several cabbage cultivars in China. During the development of dominant male sterility lines in cabbage, the conventional identification of homozygous male-sterile plants (CDMs399-3/CDMs399-3) is a laborious and time-consuming process. For marker-assisted selection (MAS) of the gene CDMs399-3 transferred into key spring cabbage line 397, expressed sequence tag-simple sequence repeats (EST-SSR) and SSR technology were used to identify markers that were linked to CDMs399-3 based on method of bulked segregant analysis (BSA). By screening a set of 978 EST-SSRs and 395 SSRs, a marker BoE332 linked to the CDMs399-3 at a distance of 3.6 cM in the genetic background of cabbage line 397 were identified. 7 homozygous male-sterile plants in population P1170 with 20 plants were obtained finally via MAS of BoE332. Thus, BoE332 will greatly facilitate the transferring of the gene CDMs399-3 into the key spring cabbage line 397 and improve the application of DGMS in cabbage hybrid breeding.     

菜心细胞质雄性不育系转育研究

以油菜波里马胞质雄性不育系为不育源,四九菜心等6个品种为轮回亲本进行杂交、回交转育研究。结果表明:经过4个世代的单株选择和回交转育,选育出菜心胞质雄性不育材料经济性状与轮回亲本相似,平均不育株率为89.4%。其中以80天特青为轮回亲本转育的不育材料表现最好,不育株率达到98%,基本达到菜心胞质雄性不育系的育种目标。     

绿宝70天菜心细胞核不育系转育研究

以法45大白菜核不育系为不育源,通过常规杂交的方法,向绿宝70天菜心转育核不育基因,初步转育成绿宝70天菜心细胞核不育系、甲型两用系和临时保持系.     

土壤盐分离子对乌鲁木齐小白菜Pb含量的影响研究

为研究土壤中盐分离子对小白菜Pb含量的影响,采取正交试验L₁₆ 4⁵和盆栽试验方法,分析了5种阳离子（Ca²⁺、Mg²⁺、K⁺、Na⁺、Pb²⁺）和3种阴离子（SO₄^2-、Cl^-、NO₃^-）对小白菜地上部和根系中Pb含量的影响。结果表明,试验条件下小白菜地上部Pb含量为0.215~0.930 mg·kg^-1,根系中Pb含量为1.648~24.33 mg·kg^-1,可食用部分超标率达81.3%。土壤盐分离子对小白菜地上部Pb含量影响顺序为： Ca²⁺ > Mg²⁺ > Na⁺ > Pb²⁺ > K⁺。根据相关性分析,土壤中Ca²⁺与小白菜地上部Pb含量呈显著负相关（r=-0.540 3,P<0.05）,Na⁺和Pb²⁺呈正相关但不显著。Mg²⁺和K⁺呈负相关,也不显著。Cl^-和NO₃^-与小白菜地上部Pb含量呈显著负相关（r=-0.540,P<0.05）,SO₄^2-呈正相关,但不显著。对小白菜根系Pb含量影响顺序为： Pb²⁺ > K⁺ > Na⁺ > Ca²⁺ > Mg²⁺,土壤中Pb²⁺对小白菜根系Pb的含量呈显著正相关（r=0.483,P<0.05）,Ca²⁺、Mg²⁺、K⁺、Na⁺、SO₄^2-、Cl^-和NO₃^-相关性不显著。乌鲁木齐土壤中固有的盐分离子对小白菜可食用部分Pb的吸收没有抑制作用。     

白菜雄性不育相关基因BcMF3功能的反义RNA验证

为了研究白菜等十字花科植物花粉发育及雄性不育发生的机理,根据编码果胶甲酯酶的白菜雄性不育相关基因BcMF3的cDNA序列设计特异引物,从普通白菜(Brassica campestris ssp.chinensis var.communis,syn.B.rapa ssp.chinensis var.communis)花蕾cDNA中扩增出458 bp的片断,构建BcMF3的反义RNA植物表达栽体,然后转化菜心(B. campestris ssp.chinensis var.parachinensis).研究发现,31.3%菜心转基因植株花粉表现出畸形,而且只有部分花粉具有活力,花粉离体萌发率降低为31.6%,其花药果胶甲酯酶活性降低了12.8%.上述结果表明BcMF3基因与普通白菜和菜心等白菜植物的花粉发育密切相关.     

寄主植物对黄曲条跳甲抗性相关酶系活性及其频率的影响

取食7种十字花科蔬菜的黄曲条跳甲成虫的酯酶(ESTs)、羧酸酯酶(CarE)和乙酰胆碱酯酶(AchE)活性及其频率分布存在不同程度的差异.取食大白菜的黄曲条跳甲成虫的酯酶平均活性最高,其次为取食菜心和上海青的,取食萝卜的最低.取食大白菜的黄曲条跳甲成虫的羧酸酯酶平均活性最高,其次为取食萝卜和菜心的,最低的是取食甘蓝的.取食不同寄主的黄曲条跳甲成虫的乙酰胆碱酯酶平均活性高低顺序按寄主排列为:萝卜>大白菜>甘蓝>上海青>山西白>黄花芥兰>菜心.     

Identification of soft rot resistance loci in Brassica rapa with SNP markers

Soft rot caused by Pectobacterium carotovorum(Pc) is a devastating disease of Brassica rapa, causing substantial reductions in crop yield and quality. Identifying genes related to soft rot resistance is the key to solving this problem.To characterize soft rot resistance, we screened a soft rot-susceptible Chinese cabbage(A03), a resistant pakchoi(‘Huaguan’), and a resistant mutant(sr). An F2 population was generated by crossing susceptible Chinese cabbage A03and resistant pakchoi ‘Huaguan’ to id...     

菜薹花药培养出胚率的影响因素研究

以红菜薹、白菜薹、菜心24个品种为试验材料,采用不同的培养基及培养方式,研究基因型、培养基成分及培养方式对不同品种菜薹花药培养诱导率的影响。结果表明:有15个品种的花药诱导出胚,各品种间的出胚率差异显著;红菜薹、白菜薹、菜心中出胚率最高的分别为H08-4(1.38胚/蕾)、B08-7(,0.757胚/蕾)、C08-4(0.682胚/蕾);采用Keller培养基+10%蔗糖+0.1%活性炭作为花药胚状体诱导培养基;花药培养出胚率以接种在双层培养基中的效果最佳。     

药用野生稻(CC)-栽培稻(AA)异源单体附加系MAAL8的遗传分析

[目的]通过对药用野生稻一栽培稻异源单体附加系MAAL8进行花药培养及其与栽培稻H1493回交,研究异源单体附加系的遗传特征。[方法]采用表型分析研究了后代分离比例,并采用SSR和甲基化分析研究了异源染色体的传递行为。[结果]在145个回交后代植株中有78株保留了MAAL8的卷叶标记性状,在32株花培植株中有5株为正常卷叶,2株为极端卷叶,其余为平叶植株。用14对SSR标记进行分析显示,在所有卷叶植株中可得到药稻特征带型,而在平叶植株中都没有。用11个多态性RFLP标记进行Methyla-tion-SensitativeSouthem分析显示,有8个在AA和CC基因组之间的甲基化变异方式是不同的。[结论]MAAL8的异源染色体可经过减数分裂完整独立地传递给后代,并保持表型特征不变,且药稻的甲基化方式随着单条染色体的附加可在杂种后代中稳定遗传,甲基化的稳定性可能对异源染色体在栽培稻基因组环境中的相对独立遗传有一定作用。     

Co-silencing E1 and its homologs in an extremely late-maturing soybean cultivar confers super-early maturity and adaptation to high-latitude short-season regions

Soybean(Glycine max(L.) Merr.), a typical short-day plant, is sensitive to photoperiod, which limits the geographical range for its cultivation. In the flowering pathway regulated by photoperiod, E1, a flowering inhibitor in soybean, plays the dominant role in flowering time regulation. Two E1 homologs, E1-like-a(E1 La) and E1-like-b(E1 Lb), play overlapping or redundant roles in conjunction with E1. In the present study, E1 and E1 La/b were simultaneously silenced via RNA interference(RNAi) in ...     

双价抗寒基因CBF3/COR15A转化大白菜的初步研究

为探索拟南芥的冷诱导转录因子CBF3和抗寒基因COR15A在"黔白"系列大白菜的转化体系,以"黔白"系列大白菜3个优良自交系作为研究对象,分别用大白菜4d苗龄的带柄子叶、半子叶、下胚轴作为外植体,利用农杆菌介导的方法将同时含有CBF3和COR15A双价抗寒基因的表达载体转入大白菜,就影响大白菜再生和遗传转化的因素进行研究。通过抗生素筛选得到大白菜T0代转基因抗性植株36株,经PCR检测后获得转基因阳性植株6株。经抗寒性初步鉴定,转基因大白菜植株抗寒性超过对照。