云南松成熟胚的体细胞胚胎的发生研究

云南松（Ｐｉｎｕｓ ｙｕｎｎａｎｅｓｉｓ Ｆｒａｎｃｈ）是我国特产的一种优良森林树种。本研究采用云南松成熟合子胚为起始外植体,在含２,４－Ｄ１０ｍｇ／Ｌ,ＫＴ和ＢＡ各４ｍｇ／Ｌ的改良Ｐ６培养基上得到胚发生培养物。将白色半透明的胚性愈伤组织（含早期原胚）在含２,４－Ｄ１．０ｍｇ／Ｌ,ＫＴ和ＢＡ各０．４ｍｇ／Ｌ的培养基上保持并增殖。在附加９,０００ｍｇ／Ｌ肌醇的高渗培养基（含ＮＡＡ０．５ｍｇ／Ｌ,ＫＴ     

菜心下胚轴原生质体培养和植株再生

以萌发３—４ 天（长约４ ｃｍ ）的菜心（Ｂｒａｓｓｉｃａ ｃａｍｐｅｓｔｒｉｓ ｖａｒ．ｐａｒａｃｈｉｎｅｓｉｓ）无菌苗苍白下胚轴为材料,酶解分离原生质体。经纯化的原生质体,在含０．５ ｍ ｇ／ＬＺＴ、０．５ ｍ ｇ／Ｌ２,４－Ｄ、１．０ ｍ ｇ／ＬＮＡＡ 和０．４ ｍ ｏｌ／Ｌ葡萄糖的Ｋ８ｐ 培养基中,进行微滴培养。在起始培养１４—１８小时,原生质体再生新的细胞壁。３６ 小时再生细胞开始第一次分裂。第三天分裂细胞频率可达３５％ 。培养第８—９ 天,可见含８—１６个细胞的小细胞团,植板率为１５％ —１８％ 。３ 周后将发育成直径为２ ｍ ｍ 的白色小愈伤组织,转到含０．３ ｍ ｇ／Ｌ ２,４－Ｄ并用ｇｅｌｒｉｔｅ半固化的培养基上,增殖成４—５ ｍ ｍ 直径的愈伤组织。在ＭＳ＋ ３．２（或１．６） ｍ ｇ／Ｌ ＢＡ＋ １．６（或０．８） ｍ ｇ／ＬＺＴ＋ ０．０１ ｍ ｇ／Ｌ ＮＡＡ＋ ０．１ ｍ ｇ／ＬＧＡ３ 和０．２％ 蔗糖的分化培养基上,获得芽的分化。切下约２ ｃｍ 长的芽苗,转移到含０．２ ｍ ｇ／ＬＩＡＡ 和２％ 蔗糖的培养基上,生根形成完整植株     

硬粒小麦单倍体原生质体培养及植株再生

由硬粒小麦（Ｔｒｉｔｉｃｕｍ ｄｕｒｕｍ Ｄｅｓｆ．）×玉米（Ｚｅａ ｍａｙｓＬ．）建立的单倍性胚性愈伤组织,在继代培养４ 个月后置于含２．０ ｍ ｇ／Ｌ２,４－Ｄ、３％ 蔗糖、２００ ｍ ｇ／Ｌ水解酪蛋白、１４６ ｍ ｇ／Ｌ谷氨酰胺和３００ ｍ ｇ／Ｌ天冬氨酸的ＭＳ液体培养基中进行悬浮培养,４ 个月后形成了生长迅速、由大小不同（０．５ ～５ ｍ ｍ ）的愈伤组织块组成的愈伤组织悬浮系。酶解试验表明,２．０％ 纤维素酶ＲＳ和０．５％ 的离析酶效果最好,而液体悬浮培养物和固体培养的愈伤组织（在酶解时用锋利的解剖刀片切成１ ｍ ｍ 左右的小块）都能释放出大量原生质体,但悬浮培养物释放出的原生质体状态较好,胞质更浓厚,用ＫＭ８ｐ 培养基以琼脂糖包埋培养方式培养时分裂频率可达５％ 左右。由原生质体再生的小愈伤组织经增殖、筛选后可获得胚性愈伤组织,将其转移至分化培养基Ⅰ（０．２ ｍ ｇ／Ｌ ２,４－Ｄ、１．０ ｍ ｇ／Ｌ ＢＡＰ、０．１ ｍ ｇ／ＬＮＡＡ、３％ 蔗糖、２００ ｍ ｇ／Ｌ 水解酪蛋白、１４６ ｍ ｇ／Ｌ谷氨酰胺和３００ ｍ ｇ／Ｌ天冬氨酸的ＭＳ固体培养基）和Ⅱ（不含２,４－Ｄ,其它成分同Ⅰ）上进行分步分化培养可再生出完整植株,分化频率约为２０％     

湿地松体细胞胚胎发生和植株再生

以湿地松的未成熟合子胚为外植体,在附加８ｍｇ／Ｌ,２,４－Ｄ和４ｍｇ／Ｌ ＢＡ的ＬＰ培养基上诱导出胚性愈伤组织。在含１ｍｇ／Ｌ,２,４－Ｄ和０．５ｍｇ／Ｌ ＢＡ的培养基上保持并增殖。提高培养基的渗透压后,愈伤组织内大量的胚性胚柄细胞团和早期原胚。     

海边香豌豆胚性愈伤组织的诱导和体细胞胚发生

将生长１４ｄ的海边香豌豆（Ｌａｔｈｙｒｕｓ ｍａｒｉｔｉｍｕｓ（Ｌ．）Ｂｉｇｅｌ）无菌苗下胚轴切成０．５ｃｍ左右的片段,置于含有１ｍｇ／Ｌ２,４－Ｄ,０．５ｍｇ／Ｌ ＢＡ和０．５％ＮａＣｌ的ＭＳ培养基中,２８ｄ后诱导出胚性愈伤组织。将其转入含有适当浓度２,４－Ｄ的ＭＳ培养基上,又２８ｄ后可得到大量球形胚和心形胚以及极少量鱼雷胚和子叶胚。诱导体细胞胚适合的２,４－Ｄ浓度为０．５ｍｇ／Ｌ。较高浓度的２     

杨树新品种叶肉原生质体培养和植株再生

从１ 个月龄的ＮＬ－８０１０６ 杨（Ｐｏｐｕｌｕｓｄｅｌｔｏｉｄｅｓ×Ｐ． ｓｉｍ ｏｎｉｉ）无菌苗叶片分离得到大量原生质体,纯化后其原生质体产量为４×１０７／ｇ ｆｒ．ｗ ｔ． 纯化的原生质体在含２,４－Ｄ ２ ｍ ｇ／Ｌ、ＮＡＡ ０．５ ｍ ｇ／Ｌ和ＫＴ ０．５ ｍ ｇ／Ｌ的ＫＭ８ｐ 和ＭＳ培养基中进行高密度液体浅层培养,渗透势为０．４０ ｍ ｏｌ／Ｌ的ＫＭ８ｐ 培养基中原生质体分裂频率最高． 培养第５ 天观察到第一次细胞分裂,培养１０ ｄ 的分裂频率为４．５％ ,１２ 周内可形成大量的细胞团和小愈伤组织． ＮＬ－８０１０６杨叶肉原生质体在富含有机氮并以葡萄糖为碳源的培养基中具有较高的分裂频率和植板率．小愈伤组织在ｇｅｌｒｉｔｅ 固化的ＮＬＺ１ 培养基上增殖生长,３ 周后形成４—６ ｍ ｍ 结构紧密的鲜红色愈伤组织,转至ＮＬＦ分化培养基,分化成苗率为１００％ ． 待芽伸长到３ ｃｍ 时,从基部切下转至１／２ ＭＳ培养基上诱导生根,形成完整植株     

粉叶小檗愈伤组织的培养及小檗碱的含量

由粉叶小檗（Ｂｅｒｂｅｒｉｓ ｐｒｕｉｎｏｓａ）的茎、腋芽、叶、实生苗的子叶及胚轴均可诱导出愈伤组织。Ｂ５＋２,４－Ｄ０．５ｍｇ／Ｌ＋ＫＴ０．２ｍｇ／Ｌ培养基对诱导较好,而Ｂ５＋２,４－Ｄ０．５ｍｇ／Ｌ＋ＫＴ０．５ｍｇ／Ｌ对愈伤组织生长较适宜。接种量在０．４－０．８ｇ（２０ｍｌ培养基）之间较为适宜。经薄层层析－分光光度法、薄层扫描法是鉴定,证明愈伤组织具有合成小檗碱的能力,含量高达１．８１４８％。     

扁蓿豆体细胞胚的诱导和植株再生

扁蓿豆实生苗的根、下胚轴、子叶、叶片和叶柄外植体,在含２,４—Ｄ２—０．２５ｍｇＬ－１与ＫＴ０．２５－２ｍｇＬ－１及２,４—Ｄ０．５ｍｇＬ－１与ＺＴ０．５ｍｇＬ－１或ＢＡＰ０．５ｍｇＬ－１与ＮＡＡ０．０５ｍｇＬ－１的ＭＳ琼脂培养基上均可产生愈伤组织．愈伤组织在含２,４—Ｄ０．５—０．１ｍｇＬ－１与ＫＴ０．５—０．１ｍｇＬ－１或ＢＡＰ０．２５＋ＮＡＡ０．０５ｍｇＬ－１的ＭＳ培养基上可诱导分化出体细胞胚．体细胞胚在无激素的培养基上发育成完整植株．用海藻酸钠包襄体细胞胚制成人工种子,其发芽率和植株转换率分别为９５％和５３％．     

在1／3海水培养基上筛选豆瓣菜耐盐变异体

系统地研究了豆瓣菜（ＮａｓｔｕｒｔｉｕｍｏｆｆｉｃａｉｎａｌｅＲ．Ｂｒ）茎段外植体对６－ＢＡ,ＮＡＡ和２,４－Ｄ的反应,确定了ＭＳ培养基附加６－ＢＡ２．０ｍｇ／Ｌ,２,４－Ｄ０．２ｍｇ／Ｌ为豆瓣菜愈伤组织诱导,继代培养基;ＭＳ培养基附加６－ＢＡ４．０ｍｇ／Ｌ为芽再生培养基;ＭＳ基本培养基为植株的生根的扦插繁殖培养基,将３２５个豆瓣菜茎切段外植体接种到含１／３海水的愈伤组织诱导培养基上,１７块外植体     

大叶紫花苜蓿愈伤组织原生质体再生植株

大叶紫花苜蓿下胚轴诱导的愈伤组织在继代培养基上生长快速,易于分散。继代第１２ｄ的愈伤组织原生质体的得率为６．５×１０７／ｇ鲜重。原生质体培养基为ＳＨ基本培养基,含有１．０ｍｇ／Ｌ２,４－０、０．５ｍｇ／ＬＢＡ、２．０ｇ／ＬＣＨ、２％蔗糖、６％葡萄糖、５ｍｍｏｌ／ＬＭＥＳ,培养密度为１．０×１０５／ｍＬ。培养至第１２ｄ时的原生质体再生细胞植板率为３．７％。由原生质体形成的小愈伤组织在含２．０ｍｇ／Ｌ２,４－Ｄ的ＭＳ固体培养基上大量增殖。增殖的愈伤组织转移至２．０ｍｇ／Ｌ２－ｉｐ＋０．１ｍｇ／ＬＮＡＡ的Ｂ５培养基上,形成体细胞胚并发育成完整植株。     

Somatic Embryogenesis and Plantlet Regeneration from Callus of Mature Zygotic Embryos of Masson Pine

Embryogenic cultures were initiated from mature zygotic embryos of masson pine (Pinus massoniana Lamb. ) on DCR medium supplemented with 2, 4-D 10 mg/L, KT and BA each at 4 mg/L. Pale and translucent calli with early stage proembryos were maintained and multiplicated on DCR medium supplemented with 2, 4-D 1.0 mg/L KT and BA each at 0.4 mg/L. Robust late-stage proembryos were obtained when the calli were cultured on DCR medium containing 9000 mg/L myo-inositol. Abscisic acid and activated charcoal promoted the formation of cotyledonary embryos at the highest frequency of 35.1 %. Mature somatic embryos could germinate and develop further into plantlets when they were isolated and cultured on a hormone-free DCR medium.     

Plant regeneration from protoplasts of soybean (Glycine max L.)

Protoplasts were isolated from immature cotyledons of six cultivars of Glycine max L. and cultured in the KP8 liquid medium supplemented with 0.2 mg/L 2,4-D, 1 mg/L NAA and 0.5 mg/L ZT. The protoplasts started to divide after 3–5 days of culture. Sustained divisions resulted in mass production of cell colonies and small calli in 6 weeks. The calli further grew to 2–3 mm on the gelritesolidified K8 medium and were transferred onto the MSB medium with 1 mg/L 2,4-D and 0.25 mg/L BA, to obtain compact and nodular calli. Shoot formation was initiated on MSB medium with 0.15 mg/L NAA, and BA, KT and ZT, 0.5 mg/L of each, with or without 500 mg/L CH. It was followed by plant regeneration. So far, 87 plants have been regenerated from 4 cultivars, and normal seeds were obtained from them after transplanting into pots.Abbreviation IAA indol-3-acetic acid - NAA naphthalene acetic acid - 2,4-D 2,4-dichlorophenoxy acetic acid - KT kinetin - BA 6-benzyladenine - ZT zeatin - CH casein hydrolysate     

中国木薯栽培种通过器官发生再生植株研究

本文研究了中国木薯栽培种四种外植体通过器官发生再生植株的条件。结果表明:在MS附加0.05mg/L TIBA,1mg/L BA的培养基上“NZ 188”初步的萌发胚状体“切头”后切口处可直接产生丛芽,出芽率为43%。“SC201”胚状体子叶块在MS附加0.5 mg/L NAA,0.5mg/L BA的培养基上可直接出芽,出芽率为42%,在MS附加0.5mg/L IBA,1.5mg/L BA培养基上·出芽率为31%,AgNO_3和ABA单独使用或配合使用均不利于芽的再生。“NZ188”胚状体下胚轴在MS附加0.5mg/LNAA,0.5mg/L BA的培养基上形成的愈伤组织转入MS附加1mg/L NAA,2mg/L BA的培养基上,3周后大多数愈伤组织有绿点出现、仅4.4%外植体分化出芽。“HZ188”无菌苗茎段接种在MS附加0.05mg/L TIBA,2mg/LBA的固体培养基上,2周后形成大量愈伤组织,4周后仅见一块愈伤组织分化出芽。     

从石刁柏原生质体培养再生植株

石刁柏,又名芦笋(Asparagus officinalisL.)是百合科天门冬属植物。其栽培品种含有丰富的维生素类及蛋白质。同时,石刁柏对于某些疾病有一定的药效,因此它已成为人们所喜爱的一种高级营养蔬菜。国外已有不少关于石刁柏试管苗繁殖的报告,但至今只有Bui Dang Ha等从石刁柏枝状叶分离的原生质体得到愈伤组织,并由此愈伤组织诱导获得了再生植株。此后,未见在石刁柏的原生质体培养方面再有新的工作。在本文中,我们利     

太白米组织培养的研究

太白米地下鳞茎在MS附加不同浓度KT、BA、IAA、NAA、2,4－D的培养基上,可诱导产生愈伤组织,其中在MS附加NAA 0.5mg/L,KT0.1mg/L的培养基上愈伤组织诱导率最高,可达65％,且生长快;培养在MS附加2,4－D 1.0mg/L,KT 0.1mg/L培养基上的鳞茎可经不定根直接发育成新的鳞茎,由此建立了太白米的鳞茎再生体系,鳞茎再生率达2．17倍。     

Plantlet regeneration from mature zygotic embryos and embryonic explants of masson pine （Pinus massoniana Lamb.）

Excised zygotic embryos,cotyledons and hypocotyls of juvenile seedlings of masson pine were grown on DCR medium supplemented with several concentrations of various plant phytohormones.BA(1.0mg/L) in combination with NAA(0.05mg/L) in DCR medium was found to increase the formation of adventitious buds from mature zygotic embryos,but most of them were formed at the tips of embryonic cotyledons.Adventitious buds were obtained from cotyledons and hypocotyls from juvenile seedlings when they were cultured on DCR medium containing BA 3-5 mg/L and NAA 0.1-0.2 mg/L.Elongation of buds were observed on hormone-free DCR medium with or without activated charcoal(0.5%).Root initiation was achieved with full or half strength DCR medium supplemented with IBA 1.0 mg/L and NAA 0.25-0.5 mg/L.Approximately 11-20 axillary buds formed on each explant when juvenile seedling explants were treated(3-20h) with BA 50-100 mg/L,followed by transfer to hormone-free DCR medium.The maximum number of shoots obtained per explant within six months was 33.     

Development of a plant regeneration system based on friable embryogenic callus in the ornamental Alstroemeria

 Stem segments of seedlings from two Alstroemeria breeding lines, cultured on media supplemented with 4 mg/l 2,4-dichlorophenoxyacetic acid and 0.5–1.0 mg/l 6-benzylaminopurine (BA), initiated soft callus, which became compact after subculture on a medium with only 0.5 mg/l BA. Friable embryogenic calli were initiated from compact callus on a medium supplemented with 10 mg/l picloram. Proembryos developed from friable embryogenic calli via embryos into plants after subculture on medium supplemented with 0.1 mg/l BA. The proembryos formed friable embryogenic calli again after culture on medium supplemented with 10 mg/l picloram. The total time needed to regenerate a complete plantlet from friable callus was approximately 6 months. This system for the production of embryogenic material is considered to have valuable applications for genetic transformation in Alstroemeria. Received: 22 April 1999 / Revision received: 16 July 1999 · Accepted: 20 July 1999     

蛇床幼茎离体培养中体细胞胚胎形成的观察

蛇床幼茎外植体经诱导产生了愈伤组织。在ＭＳ＋２,４－Ｄ,０．２ｍｇ／Ｌ＋ＺＴ０．４ｍｇ／Ｌ培养基中,愈伤组织转变成胚性愈伤组织。转入ＭＳ＋ＮＡＡ０．２ｍｇ／Ｌ＋ＺＴ０．８ｍｇ／Ｌ培养基以后,胚性愈伤组织分化出体细胞胚胎。体细胞胚胎在ＭＳ＋ＮＡＡ０．５ｍｇ／Ｌ培养基中可直接发育成为完整植析。显微观察表明,体细胞胚胎产生于愈伤组织的表层细胞或内部细胞。在鱼雷胚期已有螺纹导管的分化。子叶期的维管组织从两     

宽皮柑橘品种的胚性愈伤组织诱导

以宽皮橘3个品种的幼嫩胚珠和8个品种成熟果实中未发育胚珠为试材,采用EME(MT 500mg/L麦芽浸出物)、MKT(EME 10mg/L KT)、MGS(EME 1mg/L GA3 40mg/L硫酸腺嘌呤)和MDB (EME 0.01mg/L 2,4-D 0.1mg/L BA)4种培养基进行胚性愈伤组织诱导,结果表明EME与MKT培养基配合使用有利于从幼嫩胚珠获得胚性愈伤组织;MGS比MDB更有利于成熟果未发育胚珠胚性愈伤组织的诱导;暗培养有利于此诱导过程。经两种途径获得的胚性愈伤组织,染色体数目稳定,均为二倍体2n=2x=18。