由小-簇麦杂种悬浮细胞和原生质体再生植株

１９８８年以来,小麦原生质体培养取得了重要进展［１—７］,但成功还仅限于少数基因型,因此,为了建立和不断完善小麦及其他禾谷类植物原生质体培养的技术体系,还有待在更多的基因型中进行探索。在小麦远缘杂种系统中,１９９０年王铁邦等［８］培养小－偃麦原生质体获得成功。本文报道由普通小麦－簇毛麦杂种悬浮细胞和原生质体再生植株的结果。材料和方法起始材料取自本实验室继代保存近２年的小－簇麦杂种（２ｎ＝４ｘ＝２８）愈伤组织。该愈伤组织是由小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．,品种：“农大１４６”）×簇毛麦…     

Plant regeneration from embryogenic cell suspension cultures of wild sorghum (Sorghum dimidiatum Stapf.)

A simple and efficient protocol is described for regeneration of wild sorghum (Sorghum dimidiatum) from cell suspension cultures. Fast-growing cell suspensions were established from shoot-meristem-derived callus. Plating of the suspension on Murashige and Skoog agar medium supplemented with 2.5 mg l^–1 2,4-dichlorophenoxyacetic acid (2,4-D) resulted in the formation of embryogenic calli. High-frequency (80%) somatic embryogenesis from small cell clusters (300–400 μm) was observed when the cultures were initially maintained in liquid medium with reduced levels of 2,4-D (0.25 mg l^–1), followed by transfer to regeneration medium. Direct plating of these small clusters on regeneration medium or transfer to liquid regeneration medium containing kinetin and 6-benzylaminopurine resulted in the development of mature somatic embryos and plantlets. The regenerants developed to maturity and were all phenotypically and cytologically normal. Received: 20 May 1998 / Revision received: 1 September 1998 / Accepted: 23 September 1998     

Culture of and fertile plant regeneration from regenerable embryogenic suspension cell-derived protoplasts of wheat (Triticum aestivum L.)

Summary Regenerable embryogenic cell suspensions initiated from immature embryo-derived friable, fast growing, embryogenic calli of GK Ságvári winter wheat (Triticum aestivum L.) served as sources of protoplasts, which were cultured in different liquid or agarose-solidified media. Protocallus formation was best on KM_8p (Kao and Michayluk 1975) and GM (Li and Murai 1990) media, and protocallus growth on MS (Murashige and Skoog 1962) callus growing medium. Green shoot/plant regeneration occurred on MS regenerating medium, and rooting on MS or N6M (Mórocz et al. 1990) hormone-free media. Protocalli maintained their morphogenic capacity over 4 months, and with multiple subcultures on half-strength MS regenerating medium, the total number of regenerants could be increased. Approximately 1000 shoots/plants were regenerated and over 500 plants were transplanted in the greenhouse. The majority of them had an abnormal chromosome number and low viability, however, one plant grew to maturity and set seed.Abbreviations BAP 6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - ECS embryogenic cell suspension - GA₃ gibberellic acid - GM General medium - IAA indole-3-acetic acid - IBA indole-3-butyric acid - MS Murashige and Skoog medium - NAA 1-naphthaleneacetic acid - RECS regenerable embryogenic cell suspension     

甘薯叶柄原生质体有效植株再生

将甘薯（Ｉｐｏｍｏｅａｂａｔａｔａｓ（Ｌ．）Ｌａｍ．）‘元气’和‘白星’（‘ＷｈｉｔｅＳｔａｒ’）的叶柄原生质体培养在含有０．０５ｍｇ·Ｌ－１２,４Ｄ和０．５ｍｇ·Ｌ－１ＫＴ的改良ＭＳ液体培养基中,３～４ｄ后细胞开始分裂。培养８～９周后,将直径达１～２ｍｍ的愈伤组织转移到添加０．０５～０．２ｍｇ·Ｌ－１２,４Ｄ和０～０．５ｍｇ·Ｌ－１ＫＴ或添加０．５～２．０ｍｇ·Ｌ－１ＮＡＡ和１．０～３．０ｍｇ·Ｌ－１ＢＡＰ的ＭＳ固体增殖培养基上使其增殖。转移３～５周后,将愈伤组织再转移到ＭＳ基本培养基或转移到添加２．０～３．０ｍｇ·Ｌ－１ＢＡＰ的ＭＳ培养基上。当进一步转移到ＭＳ基本培养基上后,从愈伤组织或从愈伤组织形成的不定根上再生出植株。‘元气’植株再生率高达６０．０％,ＷｈｉｔｅＳｔａｒ高达４３．４％。     

苎麻原生质体培养及植株再生

用苎麻（Ｂｏｅｈｍ ｅｒｉａ ｎｉｖｅａ）子叶诱导愈伤组织并建立悬浮细胞系． 用４．５％ 纤维素酶、０．８％ 离析酶、０．８％ 半纤维素酶的混合酶液分离悬浮培养细胞,可得到２×１０６ 个／ｇ ｆｒ．ｗｔ的原生质体．这些原生质体以海藻酸钠包埋方式培养在附加２,４－Ｄ ０．５ ｍ ｇ／Ｌ、ＫＴ ０．５ ｍ ｇ／Ｌ 的ＫＭ８ｐ 培养基中,５０ ｄ 左右可形成肉眼可见的小愈伤组织．愈伤组织经过扩增,在不同的分化培养基上可诱导芽、根的形成,再生出完整植株． 子叶原生质体则仅能进行几次有限的分裂     

甘蔗原生质体的植株再生

从甘蔗（Ｓａｃｃｈａｒｕｍ ｏｆｆｉｃｉｎａｒｕｍ Ｌ．）嫩叶外植体诱导愈伤组织,经继代培养后,挑选胚性愈伤组织,转入ＭＳ３ 液体培养基,进行悬浮培养。当培养物分离出小粒状的细胞团,细胞变得小而圆时,用于分离原生质体。原生质体以琼脂糖固化的培养方式培养于ＭＲＰ１ 培养基中。由原生质体再生的愈伤组织有两种类型。挑选粒状、坚实的再生愈伤组织转移到Ｎ６ 分化培养基上,“新台糖１ 号”再生的愈伤组织,在含有ＫＴ ０．５ ｍ ｇ／Ｌ的培养基中,分化出绿芽并长成完整的植株。而“粤糖５７－４２３”和“ＵＳ６６－５６－９”再生的愈伤组织,在加有０．１％ 的活性炭的培养基中,前者分化出白化苗,后者分化出根     

沙打旺原生质体培养再生植株

用1%半纤维素酶,0.4%纤维素酶,0.1%果胶离析酶,CPW9M酶液分离沙打旺无菌苗下胚轴和子叶原生质体。K8P原生质体培养基悬滴培养。下胚轴原生质体形成小细胞团后用琼脂糖包埋培养,形成小块愈伤组织后转入增殖培养基M1、M2(改良MS培养基)上形成大块愈伤组织。经过两次诱导分化,在分化培养基M3(MS 0.7mg/L BA 0.2mg/L NAA),M4(MS 0.5mg/L BA 0.5mg/L KT 0.5mg/L ZT 0.2mg/L NAA)和M6(MS 3mg/L ZT 0.2mg/L IAA)上分化出苗,再生植株。由子叶分离的原生质体未能形成愈伤组织。     

籼稻原生质体培养和植株再生

用籼稻IR52、IR8和IR45的幼花序和幼胚愈伤组织在LS培养基建立了稳定的悬浮培养物。悬浮系的建立经历三个阶段:褐变期,长根期,成熟期。建立了适合籼稻原生质体生长的Y8培养基,其植板率显著高于KPR和PCM培养基。悬浮细胞系间差异明显,只有部份系可以提供有分裂能力的原生质体或具看护活性。以上三个品种的原生质体均分裂良好,但只有IR52和IR8分化出苗,其中IR52分化率1.25%,得再生植株50余株,移至田间生长结实正常。     

Plant regeneration from embryogenic suspension-derived protoplasts of ginger (Zingiber officinale Rosc.)

A procedure is described to regenerate plants from embryogenic suspension-derived protoplasts of ginger (Zingiber officinale Rosc.). Somatic embryogenic calli were induced from ginger shoot tips on solid MS medium with half the concentration of NH₄NO₃ and supplemented with 1.0 mg l⁻¹ 2,4-Dichloroacetic acid (2, 4-D) and 0.2 mg l⁻¹ Kin. Rapid-growing and well-dispersed suspension cultures were established by subculturing the embryogenic calli in the same liquid medium. Protoplasts were isolated from embryogenic suspensions with an enzyme solution composed of 4.0 mg l⁻¹ cellulase, 1.0 mg l⁻¹ macerozyme, 0.1 mg l⁻¹ pectolyase, 11% mannitol, 0.5% CaCl₂ and 0.1% 2-(N-morpholino) ethane sulphonic acid (MES) for 12–14 h at 27°C with a yield of 6.27 × 10⁶ protoplasts g⁻¹ fresh weight. The protoplasts were cultured initially in liquid MS medium with 1.0 mg l⁻¹ 2, 4-D and 0.2 mg l⁻¹ Kin. Then the protoplast-derived calli (1.5 cm²) were transferred to a basal MS medium containing 0.2 mg l⁻¹ 2, 4-D, 5.0 mg l⁻¹ benzyladenine (BA), 3% sucrose and 0.7% agar. The white somatic embryos were transferred to MS medium lacking growth regulators for shoot development. Shoots developed into complete plantlets on a solid MS medium supplemented with 2.0 mg l⁻¹ BA and 0.6 mg l⁻¹ α-Naphthaleneacetic acid (NAA). In addition, the effects of AgNO₃, activated charcoal (AC) and ascorbic acid (AA) on browning of protoplast-derived calli are discussed.     

高频水稻原生质体植株再生

在过去的几年中水稻(Oryza sativa L.)的原生质体培养取得了较大进展。我们在此基础上对国内大面积推广的优良粳稻品种的原生质体进行了高频植株再生的实验,并对两种不同的培养方法——琼脂糖包埋法和看护培养法进行了初步的比较研究。     

霞草原生质体培养和植株再生

用霞草胚性悬浮细胞分离原生质体,以含0.2%琼脂糖的KM 8p培养基薄层漂浮培养,原生质体培养密度6×10~3-1×10~4/ml。培养3天再生细胞开始分裂,7天统计分裂频率最高达25.4%,10天形成小细胞团,并加降低渗透压的稀释培养基,每周一次。20—25天形成肉眼可见的小愈伤组织,植板率达3.5%。原生质体衍生的愈伤组织在增殖培养时加入0.3%-0.4%活性炭有利于生长及分化。在含6-BA 3.5 mg/L,IBA 0.8 mg/L的培养基上,再生芽的分化频率可达85%。再生芽在添加NAA 0.5 mg/L,6-BA 0.05 mg/L的1/2 MS生根培养基中2周内形成具根的再生小植株。     

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