Cytogenetic analysis of F1, F2 and BC1 plants from intergeneric sexual hybridization between Sinapis alba and Brassica oleracea by genomic in situ hybridization

By intergeneric sexual hybridization between Sinapis alba and Brassica oleracea , F₁, F₂ and BC₁ progeny plants were produced. S. alba plants (genome SS, 2n = 24) were pollinated with B. oleracea (genome CC, 2n = 18), and the fertile F₁ plants were pollinated with B. oleracea to obtain BC₁ plants. GISH analysis showed that 10 out of 12 F₁ plants had 12 S. alba chromosomes (one full S chromosome set) and nine B. oleracea chromosomes (one C chromosome sets), representing the expected hybrids. However, two F₁ plants had 12 S chromosomes and 18 C chromosomes (two C chromosome sets), indicating unexpected hybrids. A maximum of three trivalents between C and S chromosomes were identified at metaphase I of semi-fertile F₁ pollen mother cells (PMCs), which indicates homology and chromosome pairing between these two genomes. The C genome had obviously been doubled in two F₂ plants from selfed semi-fertile F₁ plants. BC₁ plants consisted of 18 C chromosomes and different numbers of one, five and six additional S chromosomes, respectively. Monosomic alien addition lines developed in the present study can be used for B. oleracea breeding and Sinapis alba gene mapping.     

平菇汁对野蘑菇生长的研究

野蘑菇具有很高的营养和药用价值,但人工培养时生长缓慢、老化快、产量低,使得在菌剂等实际生产应用方面受到限制。结合有关文献资料,发现平菇汁中有不知名组分对野蘑菇菌丝生长有显著的促进作用,旨在找出并确定这种物质及其最适生长浓度。分别以水、乙醇、丙酮为透析液, MWCO为8000~14000的透析袋透析等体积蘑菇汁, DNS法测还原糖、双缩脲法测蛋白质、酸性染料比色法测生物碱、苯酚-硫酸法测总糖、甲醛法测氨为测定手段,菌丝直径为测量指标。蛋白质多的组分加入培养基中野蘑菇菌丝生长迅速,生物碱多的组分加入培养基野蘑菇菌丝生长缓慢。平菇汁中有类蛋白物质起促进野蘑菇菌丝生长作用,生物碱类物质起抑制野蘑菇菌丝生长作用。     

属间体细胞杂交创建甘蓝型油菜细胞质雄性不育系及其鉴定

 利用原生质体融合技,获得了甘蓝型油菜品种中双4号与新疆野生油菜野油18的对称性体细胞杂种。杂种当代建成的6株植株中有2株雄性不育株。用中双4号做轮回亲本与不育株回交,不育株率随回交代数增加而升高,至BC3代大部分株系的不育株率接近100%,到BC4代不育性稳定,群体中无育性分离现象。用波里马CMS的恢复系和保持系与其测交,6个甘蓝型油菜品系均可保持该不育系的不育性,表明该雄性不育胞质与Pol不育胞质不同。     

环境因素对新疆不同地区田旋花种子萌发的影响

【目的】研究环境因素对新疆不同地区田旋花种子萌发的影响,为制定有效的杂草防除技术提供理论依据。【方法】通过室内测定,研究温度、盐碱度、干旱、pH值等环境因子对新疆阿拉尔市和新和县两个点田旋花种子萌发的影响。【结果】阿拉尔市、新和县田旋花种子在5～50℃恒温范围内,随着温度的升高发芽率先升高后降低;田旋花种子最低发芽温度为5℃,当温度上升到30～40℃,两个县市的田旋花种子发芽率均达到最大值,种子在15℃/5℃～45℃/35℃的变温条件下均可发芽,且当温度为40℃/30℃时发芽率最高。盐碱胁迫可降低田旋花种子的发芽率,当NaCl浓度大于300 mmol/L或NaHCO₃浓度大于250 mmol/L时,两个县市的田旋花种子不能萌发;同时,田旋花种子的萌发受水势的影响较大,随着水势的增加,田旋花种子的发芽率逐渐降低,当水势达-0.6 MPa时,新和县田旋花种子已不能萌发,当水势达到-1.0 MPa时,阿拉尔市田旋花种子的萌发率仅为2%;pH值5～10,两个县市的田旋花种子均能正常萌发。【结论】室内模拟实验条件下,温度、盐碱和水势均是影响新疆不同地区田旋花种子萌发的关键环境因素,而农田生态系统中,干旱胁迫和pH不是限制田旋花萌发的主要因素。     

侵染苣荬菜的中国胜红蓟黄脉病毒及伴随的卫星基因组结构特征

杂草是菜豆金色花叶病毒属病毒的重要中间寄主,常富集多种该属病毒及其卫星病毒。2014年,在云南红河常见杂草苣荬菜Sonchus arvensis上出现了疑似菜豆金色花叶病毒属病毒病症状。利用克隆、测序和生物信息学分析技术对其所含病毒进行分离鉴定,结果从1株病样中共获得了两条菜豆金色花叶病毒属病毒全序列、两条β卫星全序列和一条α卫星全序列。序列分析显示,两条菜豆金色花叶病毒属病毒全序列与中国胜红蓟黄脉病毒相似性最高,分别为99%和96%,确定为中国胜红蓟黄脉病毒的分离物。两条β卫星全序列与赛葵黄脉β卫星相似性最高,为97%,确定为赛葵黄脉β卫星的一个分离物。α卫星全序列与中国番茄黄化曲叶α卫星相似性最高,为86.3%,是中国番茄黄化曲叶α卫星的一个分离物。这是菜豆金色花叶病毒属病毒病害复合体在中国侵染苣荬菜的首次报道。     

Plant regeneration from in vitro cultures of cotyledon explants and anthers of Sinapis alba and its implications on breeding of crucifers

Summary Protocols for plant regeneration from cotyledon explant and anther cultures of Sinapis alba have been developed for creating doubled-haploids and somaclonal variation. Among the several cultivars tested in this study, only Arda responded well to in vitro plant regeneration both from anther-as well as cotyledoncultures. Multiple shoot formation in cotyledon explants, which always followed a brief callusing phase, was found to be the best on MS medium with ZEA (1.0mg/l) and NAA (0.1mg/l). Regeneration frequency declined sharply in the absence of auxin or presence of other cytokinins and/or auxin. The frequency of shoot regeneration also declined with reduction in the photoperiod to 16h. On MS + BAP (1.0mg/l) + NAA (1.0mg/l) medium, cotyledonary explants showed profuse callusing, which could regenerate shoots on high ZEA + low NAA/IAA medium. However, it declined with progressing time in culture. Anthers, excised from fresh as well as cold pretreated buds, cultured on 10% sucrose containing MS media with different hormonal constitution, developed calli and/or embryos. Initial culture temperature was important with embryogenesis occurring only in anthers cultured at 30°C for 3 weeks. A high temperature (35°C) treatment was lethal for both callus as well as embryo formation. While BAP + NAA and ZEA + NAA/IAA supported embryogenesis, further plant regeneration from anther-or embryo-callus could be achieved in ZEA + NAA/IAA media. Some of the regenerants flowered already in vitro and had small and sterile flowers. Cytological examination of some of the root differentiating calli indicated the presence of haploid as well as diploid cells. Shoots were rooted during prolonged incubation on the same medium or on transfer to MS (reduced)/ B5 + ZEA + NAA media.     

Production and characterization of intergeneric hybrids between Brassica oleracea and a wild relative Moricandia arvensis

Intergeneric crosses were made between Brassica oleracea and Moricandia arvensis utilizing embryo rescue. Six F₁ hybrid plants were generated in the cross‐combination of B. oleracea × M. arvensis from 64 pods by the placenta‐embryo culture technique, whereas three plants were produced in the reciprocal cross from 40 pods by the ovary culture technique. The hybrid plants were ascertained to be amphihaploid with 2n = 23 chromosomes in mitosis and a meiotic chromosome association of (0–3)II + (17–23)I at metaphase I (M I). In the backcross with B. oleracea, some of these hybrids developed sesquidiploid BC₁ plants with 2n = 32 chromosomes that predominantly exhibited a meiotic configuration of (9II + 14I) in pollen mother cells. The following backcross of BC₂ plants to B. oleracea generated 48 BC₃ progeny with somatic chromosomes from 2n = 19 to 2n = 41. The 2n = 19 plants showed a chromosomal association type of (9II + 1I) and a chromosomal distribution type of (9¹/₂ + 9¹/₂) or (9 + 10) at M I and M II, respectively. These facts might suggest that they were monosomic addition lines (MALs) of B. oleracea carrying a single chromosome of M. arvensis that could offer potential for future genetic and breeding research, together with other novel hybrid progeny developed in this intergeneric hybridization.     

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.