Cloning and Characterization of the Microspore Development-Related Gene BcMF2 in Chinese Cabbage Pak-Choi （Brassica campestris L. ssp. chinensis Makino）

For the sake of providing some important information relevant to the study of the molecular mechanism of genic male sterility in plants, gene differential expression in flower buds at different developmental stages, as well as in rosette leaves, florescence leaves, and scapes was analyzed using cDNA amplified fragment length polymorphism (cDNA-AFLP) in the genic male sterile A and fertile B line of Chinese cabbage pak-choi. Following amplification of 125 pairs of primer combinations, 11 differential fragments were obtained, of which eight were from the B line and the other three were from the A line. Of 11 differential fragments, four were verified by Northern hybridization that were expressed preferentially in fertile flower buds. Results of GenBank BLAST showed that one fragment was with unknown function, whereas the other fragments have strong nucleotide sequence similarities with the polygalacturonase (PG) gene, the pectinesterase (PE) gene, and the polygalacturonase inhibitory protein (PGIP4) gene. Only fulllength cDNA from the differential fragment BcMF-A 18T 16-1 was amplified by rapid amplification of cDNA ends (RACE) and Northern analysis showed that this fragment was expressed only in medium and largesized flower buds of the B line. The full-length cDNA, designated as BcMF2 (Brassica campestris Male Fertile 2), was 1 485 bp long and was composed ofa 1 263-bp open reading frame, which had 83% nucleotide similarity to a PG gene from Arabidopsis encoding polygalacturonase. Analysis of the basic structure of the protein revealed that it had one polygalacturonase active site (RVTCGPGHGLSVGS) at 256th site of amino acids and was classified as being a member of family 28 of the glycosyl hydrolases. The role of the BcMF2gene on microspore development is discussed in the present paper.     

Promotion by 5-Aminolevulinic Acid of Germination of Pakchoi (Brassica campestris ssp. chinensis var. communis Tsen et Lee) Seeds Under Salt Stress

The seed germination and seedling growth of pakchoi (Brassica campestris ssp. chinensis var.communis Tsen et Lee cv. Hanxiao) were not significantly inhibited until the concentration of NaCl was increased to150 mmol/L. Treatment of pakchoi seeds with exogenous 5-aminolevulinic acid (ALA), at concentrations ranging from 0.01 to 10.00 mg/L, promoted seed germination when seeds were stressed by salinity, whereas levulinic acid (LA), an inhibitor of ALA dehydrase, significantly inhibited seed germination and seedling growth, suggesting that metabolism of ALA into porphyrin compounds was necessary for seed germination and seedling growth. Determination of respiratory rate during seed germination showed that ALA increased seed respiration under both normal conditions and salt stress. Furthermore, salt stress decreased levels of endogenous ALA, as well as heme, in etiolated seedlings. More salt-tolerant cultivars of pakchoi contained higher relative levels of endogenous ALA and heme under conditions of salt stress.These results indicate that salt stress may inhibit the biosynthesis of endogenous ALA and then heme,which is necessary for seed germination, and treatment of seeds with exogenous ALA prior to germination may be associated with the biosynthesis of heme.     

Successful application of microspore culture in pakchoi (Brassica campestris L. ssp. chinensis Makino var. communis Tsen et Lee) for hybrid breeding

Protoplasma - Microspore embryogenesis is an effective method of obtaining double haploid (DH) lines in only 1 year. However, the microspore embryogenesis protocol was not efficient in...     

水杨酸对小白菜抗铅胁迫的诱导

0.3 mmol/L铅胁迫下小白菜（Brassica rapa var. chinensis）生长受到显著抑制,0.1 mmol/L水杨酸（salicylic acid）诱导处理能显著缓解0.3 mmol/L铅胁迫对小白菜幼苗生长的抑制;并且水杨酸对铅胁迫下小白菜的生长存在“低促高抑”效应,当水杨酸处理浓度达到1.5 mmol/L时,不但不能缓解铅对小白菜的毒害,反而加重毒害,这可能与高浓度水杨酸与铅形成的复合伤害有关。     

菜心耐羟脯氨酸初选系的耐热性

分析了离体筛选出的一个菜心耐羟脯氨酸选择系的脯氨酸含量和耐热性。Hypr01比原品种六十天特青有较高的游离脯氨酸含量：35℃热胁迫下Hypr01中有显著的游离脯氨酸含量增加,为六十天特青的4倍和耐热品种四九菜心的近2倍。与对照相比,在35℃下连续培养的Hypr01愈伤组织表现出更强的活力和根的分化能力,鲜重增量明显加大：相对电解质渗出率、愈伤组织褐变程度和MDA含量低：SOD、CAT活性较高。在人工气候箱模拟栽培高温逆境下,Hypr01再生苗生长优于未经选择的六十天特青,并具较低的电解质渗出率和较高的鲜重增长,说明有较强的耐热性。     

外源赤霉素GA3对混合盐碱胁迫下小白菜种子萌发的影响

以小白菜（Brassica campestris ssp．chinensis L．）为材料,通过采用不同浓度的外源赤霉素（GA3）浸种处理,研究了GA3对混合盐碱胁迫下小白菜种子的发芽率（Gr）、发芽势（Gv）、发芽指数（Gi）以及根毛区宽度和胚长的影响。结果表明：混合盐碱胁迫下,小白菜各发芽指标均显著低于对照,GA,处理后,各指标均显著提高,且随外源GA3浓度的增加,各指标均表现出先增后减的趋势,当GA3浓度达80mg／L时,各发芽指标达到峰值,说明80mg／LGA3可使混合盐胁迫下小白菜种子萌发的缓解作用达到最佳。同时外源GA3处理还可不同程度地缓解盐胁迫对小白菜种子a-淀粉酶活性的抑制。     

白菜细胞核雄性不育两用系的细胞学观察

对白菜细胞核雄性不育两用系进行了花粉母细胞减数分裂和小孢子发育的细胞学观察,实验结果初步表明不育系小孢子败育时期在减数分裂末期Ⅱ这一阶段,败育方式是不能形成四分体,随后小孢子内颗粒状的内含物不断外溢,直至成为一个空壳,药室萎缩,导致花粉败育。     

白菜小孢子发育相关基因BcMF3的分离及序列分析

利用cDNA-AFLP技术对‘矮脚黄’白菜(Brassica campestris L．ssp．chinensis Makinovar．communis Tsen et Lee cv,Aijiaohuang)雄性不育两系不育株系与可育株系的表达差异进行分析,结果以A18和T16引物为引物对在可育的中蕾和大蕾中筛选出一条474bp特异表达的差异条带BA18-T16,配合RACE方法扩增了该片段的3’和5’末端,获得了该基因的cDNA全长序列BcMF3。BcMF3全长2082bp,开放阅读框长度为1755bp,编码584个氨基酸。序列分析结果表明,BcMF3基因与油菜、大白菜和拟南芥的花药特异表达果胶甲酯酶(PME)基因具有较高的相似性,由此推定,BcMF3基因为白菜花药特异表达的果胶甲酯酶基因。     

水杨酸调控盐胁迫下羽衣甘蓝种子萌发的机理

盐胁迫是植物种子萌发与植株生长的重要限制因子。以羽衣甘蓝(Brassica oleracea var.acephala)名古屋为材料,研究不同盐分对其种子萌发的影响,探索水杨酸(SA)及其合成抑制剂氨基茚磷酸(AIP)处理对羽衣甘蓝种子萌发的调控效应。实验结果表明,150与200 mmol·L^–1 NaCl处理后的羽衣甘蓝种子活力显著降低。盐胁迫显著降低种子的吸水速率、种子活力与幼苗质量,降低苯丙氨酸裂解酶活性与内源SA含量,提高过氧化氢(H2O2)与超氧阴离子(O2^–.)含量。SA可以缓解盐胁迫对羽衣甘蓝种子活力的抑制作用,通过促进内源SA合成,从而提高种子吸水率与种子活力,促进种子对K^+、Mg^2+的吸收,降低Na+含量。此外,外源施加SA能够显著增强超氧化物歧化酶和过氧化物酶活性,降低H2O2与O2^–.的积累。相反,氨基茚磷酸(AIP)处理能够增强盐胁迫对种子萌发的抑制作用,推测这与AIP处理能够显著降低种子内源SA含量密切相关。研究表明外源SA主要通过提高保护酶活性、降低活性氧积累和维持体内离子平衡来增强羽衣甘蓝的耐盐性。     

外源5-氨基乙酰丙酸对盐胁迫下菘蓝种子萌发及幼苗抗氧化酶活性的影响

采用砂培方式,研究了外源5-氨基乙酰丙酸(ALA)对盐胁迫下菘蓝种子的萌发、幼苗叶片的可溶性糖含量、丙二醛(MDA)含量及其抗氧化酶活性的影响,探讨ALA缓解菘蓝受盐胁迫伤害的响应机制。结果显示:(1)菘蓝种子萌发及幼苗生长在100 mmol·L^-1 NaCl胁迫下受到明显的抑制,种子发芽率、发芽势、发芽指数、活力指数与自然含水量均显著降低,丙二醛含量、可溶性糖含量以及超氧化物歧化酶(SOD)、过氧化物酶(POD)活性显著升高。(2)盐胁迫下适宜浓度的ALA处理显著提高了种子萌发率、自然含水量及SOD、POD和CAT活性,降低了可溶性糖和丙二醛的含量,并以16.7 mg·L^-1 ALA处理盐胁迫下菘蓝种子的发芽率、发芽势最大,其幼苗的SOD、POD、CAT活性最强。研究表明,盐胁迫显著抑制菘蓝种子的萌发及幼苗生长,适宜浓度的ALA能够有效缓解盐胁迫对菘蓝种子萌发及幼苗生长的伤害,提高植株的抗盐性,并以16.7 mg·L^-1 ALA处理效果最佳。     

红菜薹雄性不育系花药败育的细胞形态学观察

采用石蜡切片技术,在光学显微镜下系统研究了红菜薹(Brassica campestris L.ssp．chinensis L.var．utilis TsenetLee．)波里马胞质雄性不育系(Polima CMS)、红菜薹萝卜胞质雄性不育系(Ogura CMS)及相应保持系花药发育过程的细胞形态学特征。观察结果表明：红菜薹Polima CMS花药发育受阻于孢原细胞阶段,不形成花粉,属无花粉型,此不育系花药不形成绒毡层和中层;而红菜薹Ogura CMS花药败育发生于小孢子母细胞期或四分体时期,表现为绒毡层细胞异常,挤压四分体,导致四分体和绒毡层同时解体而败育。