棉花川239体胚发生和植株再生

以长江流域棉花新品系川２３９为材料,系统比较了四种激素组合（２,４－ Ｄ＋ ＫＴ,ＮＡＡ＋ ＫＴ,ＩＡＡ＋ ＫＴ, ＺＴ）对愈伤组织诱导和体胚发生的影响,对获得大量再生植株的条件进行了研究,获得了“川２３９”再生植株。并对难于进行体胚发生的棉花品种的体细胞培养策略进行了初步探讨     

Effect of incubation temperature regimes and culture medium on broccoli microspore culture embryogenesis

Conditions for reliable induction of embryogenesis from isolated microspores were studied in ten genotypes of broccoli. Embryo yields were significantly increased in almost all of the broccoli genotypes by the incubation at 32.5 ^°C for 1 day, than when the standard incubation at 30 ^°C for 2 days was used. Treatments of 48 hours at 32.5 ^°C produced less than optimal results suggesting that broccoli microspores are more sensitive to high temperatures than those of B. napus. The use of the ¹/₂ NLN-13 medium yielded greater number of embryos than the standard NLN-13. The magnitude of the response to the redution of the concentration of major salts by half in the NLN medium varied with the different genotypes. High embryogenic broccoli cultivars, such as ‘Shogun’, ‘SDB9’, and ‘Green Valiant’, presented a better response to the reduction of the concentration of major salts by half in NLN-13. Reduction never produced a detrimental effect on embryo yield and seems not to have any effect in the subsequent development of embryos in plants. This revised version was published online in July 2006 with corrections to the Cover Date.     

A Simple Method of Inducing Somatic Embryogenesis in Brassica juncea (L.) Czern & Coss.

A single-step method for the induction and development of somatic embryoids from hypocotyls explains of Brassica juncea is reported. On modified MS medium containing 2 % sucrose, 0.25 mgl ¹ 2,4-D, 0.5 mgl ¹ each of NAA and BaP-R, each explant calluses at both of and at its best, 31% of explants produce embryoids. In the variety RLM-198, the number of embryoids ranges from 8–21 per culture. Each embryoid, upon proliferation, developed up to the 25 shoots. The method is rapid; the time La ken from inoculation to the development of intact plantlets is 8–10 weeks. Regenerated plants have flowered normally and have set seed. The system can profitably be used for in vitro mutant selection and early bulking in mustard.     

The Effect of Carbohydrate Composition and Concentration on Anther Culture Response in Barley (Hordeum vulgare L.)

Culture medium composition is critical for the successful induction of microspore division in vitro. The present experiments have focused on a relatively neglected area of cell and tissue culture research, namely the carbohydrate component used in the medium. Three spring barley genotypes were cultured on a medium which was modified by replacing sucrose with the following carbohydrates (6 % w/v): maltose, fructose, malt extract, galactose and a glucose (3 % w/v)/fructose (3 % w/v) mixture. Both maltose and malt extract were superior to sucrose in their capacity to induce green plantlet differentiation from microspores. The concentrations of both sucrose and maltose were also varied. Overall the response of anthers on maltose based media was higher than on sucrose based media. Furthermore, a concentration of maltose m the range 6—12 % w/v produced a higher frequency of green plants than a low concentration (1—3 % w/v). The effect of maltose based media on germplasm of direct relevance to barley breeders was also tested. The cultivar ‘Blenheim’ was shown to be very responsive and this genetic factor was transmitted to the F₁ hybrid. The frequency of haploid to diploid regenerants was not consistent over genotypes, but in general there were more haploid than diploid regenerants. The implications of these results for barley breeding are discussed.     

蔗糖对水曲柳体细胞胚发生的影响

以水曲柳子叶为外植体,研究了不同浓度的蔗糖对水曲柳愈伤组织和体细胞胚发生以及体细胞胚数量、质量的影响。结果表明:当NAA多于BA时,附加7%蔗糖的体细胞胚发生率最高;当BA多于NAA时,附加10%蔗糖的体细胞胚发生率最高,但每个外植体上体细胞胚的数量和质量不是最好的。综合认为,选定9%~12%的蔗糖浓度最有利于水曲柳体细胞胚的诱导。     

miR396在落叶松体细胞胚胎中的功能研究

[目的]探讨miR396在落叶松体细胞胚胎发生中的功能.以期该研究能为解决体胚发育中子叶畸形胚与生根率低的问题提供新的视角,同时也为进一步揭示体胚发育的分子调控网络做基础铺垫.[方法]构建pre-miR396超表达载体,通过农杆菌介导侵染,将pre-miR396转入落叶松胚性细胞中,筛选稳定抗性细胞系.在DNA与RNA水平上,对抗性细胞系进行的鉴定、检测,同时统计比对其与野生细胞系间发芽率、叶片数目以及生根率的差异.[结果]在抗性细胞系基因组中,扩增出HTP与miR396的特异性片段,且无农杆菌Virg基因片段.RNA表达水平上,抗性系pre-miR396与miR396表达皆增加,而靶基因LaGRFs的表达量下调.统计表明抗性系胚的发芽率、生根率、叶片数目畸形率分别为70.60%、0.60%、37%,而野生型分别为92.50%、10.55%、4%,抗性系胚的发芽率、生根率显著降低(Sig.=0.000),叶片数目畸形率显著增加(Sig.=0.000).[结论]miR396在落叶松体细胞胚胎发生过程中,可能通过负调节靶基因LaGRFs的表达或者通过LaGRFs影响与它相互作用的基因,参与体胚子叶原基细胞代谢,从而影响了子叶与叶片的发育;通过负调节LaGRFs或者其它与根部发育相关的靶基因,影响了根端分生区细胞的活动,参与调控根的发育.     

云杉属树种体细胞胚胎发生研究进展

云杉属树种具有适应性强、抗逆性强、生长迅速等特性,是园林绿化的重要树种。体细胞胚胎发生技术在云杉属树种中的应用,为其品种改良和规模化、产业化繁殖提供了可能。文中综述云杉属树种体细胞胚胎发生的过程及主要影响因素,提出在云杉属树种体细胞胚胎诱导过程中存在的问题和研究展望,以期为云杉属树种体细胞胚产量和质量的提高以及建立高效再生体系提供借鉴。     

Cryopreservation of yellow-poplar and sweetgum embryogenic cultures

Cryopreservation has become anessential tool for operational application offorest tree embryogenic cultures, due to thelong evaluation periods needed for treesregenerated from these cultures. Fiveyellow-poplar (Liriodendron tulipifera)and seven sweetgum (Liquidambar spp.)embryogenic culture lines werestored in liquid nitrogen for 48 hours, afterwhich they were thawed and tested for regrowthand ability to produce somatic seedlings.Combinations of two sorbitol pretreatments andthree dimethylsulfoxide (DMSO) cryoprotectantlevels were evaluated for their impact onrecovery following cryogenic storage. The bestresults were obtained with 0.4 M sorbitol and5% DMSO, which provided 100% recovery.Somatic seedlings were regenerated from allculture lines and treatments, except for atransgenic sweetgum line.     

花楸树体细胞胚与合子胚的发生发育

利用花楸树人工控制授粉后的未成熟合子胚进行体细胞胚的诱导培养,通过石蜡切片、扫描电镜和透射电镜对合子胚和体细胞胚的发生发育过程进行组织、细胞观察。结果表明:1)体细胞胚发生进程与合子胚相似,均经历了球形期、心形期、鱼雷形期,最终发育成成熟胚;2)直接发生和间接发生在愈伤组织表面的体细胞胚均为单细胞起源,间接发生在愈伤组织内部的体细胞胚存在单细胞起源,同时还存在可能的多细胞起源;3)子叶和愈伤组织表面发生的体细胞胚在发育早期均具明显的胚柄,鱼雷形胚期胚柄退化;4)愈伤组织内部发生的体细胞胚没有胚柄;5)与合子不同,胚性细胞的第1次分裂为均等分裂;6)多细胞原胚周围降解的细胞具有细胞程序性死亡的形态特征。     

红豆杉及南方红豆杉体细胞胚胎发生的研究

利用江西的红豆杉（Ｔａｘｕｓｃｈｉｎｅｎｓｉｓ）、福建的南方红豆杉（Ｔａｘｕｓｃｈｉｎｅｎｓｉｓｖａｒ．ｍａｉｒｅｉ）以及四川的南方红豆杉（Ｔａｘｕｓｃｈｉｎｅｎｓｉｓｖａｒｍａｉｒｅｉ）的成熟胚为试验材料,成功地诱导出大量的愈伤组织。这３种材料愈伤组织的最高诱导频率分别为９５７％、９８４％、９１７％。细胞学和形态学的观察结果表明：在适当的条件下经过一段时间的培养,这些愈伤组织能够产生处在不同发育阶段的体细胞胚。上述３种植物材料每１００块愈伤组织平均最多分别可产生１２７、９１和３３个体细胞胚。