二棱大麦体细胞胚性无性系的建立

对１２个品种的二棱大麦（Ｈｏｒｄｅｕｍｄｉｓｔｉｃｈｕｍ２ｎ＝２４）幼穗和６个品种的幼胚在附加５００ｍｇ／Ｌ酪蛋白,１５０ｍｇ／Ｌ天门冬素和２ｍｇ／Ｌ２,４－Ｄ的ＭＳ培养基（Ｍ＿（３２０））中进行了离体培养,并诱导产生出了愈伤组织。把由Ｄ＿１幼胚所获得的愈伤组织转移到含２,４－Ｄ０．５ｍｇ／Ｌ（Ｍ＿（３０５））培养基中后诱导产生出了体细胞胚性愈伤组织,幼苗和根。筛选出了体细胞胚性无性系。在长达二年的继代培养中,该细胞系仍保持旺盛的再生和分化成苗能力。     

Effects of abscisic acid and proline on adaptation of tobacco callus cultures to salinity and osmotic shock

Tobacco callus ( Nicotina tabacum cv. Badischer Geudertheimer) took up sorbitol rapidly and without a lag period from media with up to 0.7 M of the polyol. Accumulation of proline was greatly enhanced under these conditions and was proportional to the absorbed sorbitol, while the viability of the callus cultures was quite low after a few hours of incubation. Under moderate conditions (0.1 M sorbitol) as well as under severe osmotic shock (0.7 M sorbitol), the cells adapted by adjusting the sorbitol/proline ratio to ca 3. NaCl (0.1 M ) had the same effect as sorbitol (0.7 M ) on the survival rate, but only slightly affected proline synthesis in the first hours of incubation. Addition of 10⁷ or 10 ⁵ M abscisic acid (ABA) did not increase the proline content, but 10 ⁷ M ABA delayed the deleterious effect of NaCl and improved the state of the cells. No influence of abscisic acid during the incubation with sorbitol could be detected. Two different strategies for the adjustment of tobacco callus to salinity and sorbitol are suggested: Non-ionic stress is controlled by the accumulation of proline, whereas ABA could be involved in the adaptation to ionic stress.     

Organogenesis and a general procedure for plant regeneration from callus culture of a commercial duboisia hybrid (D. leichhardtii x D. myoporoides)

Summary The Duboisia hybrid (D. leichhardtii x D. myoporoides) was regenerated from callus derived from shoot tips and young seeds on Murashige and Skoog (MS) medium containing 54M 1-naphthalenacetic acid (NAA) and 1M 6-benzylaminopurine (BA). Callus derived from young seeds was superior to shoot tip callus. Buds were induced from callus on MS medium supplemented with 22M BA. The buds formed shoots when transferred to MS medium with 5 M BA and 0.5M NAA. Shoots were induced to form roots when placed on MS medium containing 25 M indolebutyric acid (IBA). Plantlets so formed were transplanted and subsequently planted out in the open. Approximately 6 months were required for the full regenerative process. Trees so formed have been grown for 3 years, reached a height of 3 metres and flowered and fruited normally.     

Growth, proline accumulation and ion content in sodium chloride-stressed callus of date palm

Summary Growth and physiological responses of date palm. Phoenix dactylifera L. cv. Barhee, callus to salinity stress were examined. Callus induced from shoot tips of offshoots was cultured on Murashige and Skoog medium supplemented with NaCl at concentrations ranging from 0 to 225 mM, in consective increments of 25 mM. Data obtained after 6 wk of exposure to salt have shown a significant increase in callus proliferation in response to 25 mM NaCl the lowest level tested, beyond which callus weight decreased. At 125 mM NaCl and higher, callus growth was nearly completely inhibited. Physiological studies on callus exposed to salt stress have shown an increase in proline accumulation in response to increased salinity. Proline accumulation was correlated to callus growth inhibition. Furthermore, increasing the concentration of NaCl in the culture medium generally resulted in a steady increase in Na⁺ and reduction in K⁺ concentrations. However, at 25 mM NaCl, the only level at which callus growth was significantly enhanced, an increase in K⁺ content was noted, in comparison to the NaCl free control. In response to increasing external NaCl level, the Na⁺/K⁺ ratio increased The Na⁺/K⁺ ratio was positively correlated to proline accumulation and hence callus growth inhibition. This study provides, an understanding of the response of date palm callus to salinity, which is important for future studies aimed at developing strategies for selecting and characterizing somaclonal variants tolerant to salt stress.     

木那格葡萄快速繁殖及愈伤组织的诱导和继代

研究了木那格葡萄快速繁殖并确定了木那格葡萄愈伤组织诱导和继代的适宜条件。将木那格葡萄种子无菌处理,获得无菌幼苗,以单芽茎段为外植体,对木那格葡萄快速繁殖条件进行优化;以叶片为外植体,确定愈伤组织诱导和继代的适宜条件。结果表明：使用70%乙醇灭菌30s,0．1%升汞灭菌3min,使用浸湿的滤纸培养,适宜木那格葡萄种子萌发,萌发率为70%;适宜单芽茎段不定芽再生的培养基为B5 6-BA 0．05mg/L IBA 0．3mg/L 蔗糖30g/L,不定芽增殖倍数为1．25;适宜不定芽生根的培养基为B5 IBA 0．2mg/L 蔗糖30g/L,平均单芽生根数为5．09,平均根长2．97cm;适宜愈伤诱导的培养基为B5 6-BA1．5mg/ L NAA 0．2mg/L 蔗糖30g/L,愈伤诱导率为100%;适宜愈伤继代的培养基为B5 6-BA 1mg/L 2,4-D 0．1mg/L 蔗糖30g/L。     

Rapid mass propagation of Tylophora indica Merrill via leaf callus culture

An efficient protocol has been developed for rapid mass propagation of Tylophora indica from leaf derived callus. Optimal callus was developed from leaf explants on Murashige and Skoog (MS) basal medium supplemented with 10 2,4,5-T. Adventitious shoots were regenerated (85%) from the surface of the callus on MS medium supplemented with 5 M Kinetin. Individual elongated shoots were rooted on half-strength MS medium containing 0.5 M IBA. Regenerated plantlets with well developed shoots and roots were successfully transferred to soil. The study demonstrated a dedifferentiated callogenic propagation route via adventitious shoot development in T. indica, which could be useful for large scale multiplication of this endangered medicinal plant.     

胡杨愈伤组织质膜的两相分离法及其H~+-ATPase的特性

以胡杨愈伤组织为材料,用PEG3350/DextranT500构成的两相系统提取质膜微囊,研究质膜H+-AT-Pase的特性。结果显示:由6.3%PEG3350、6.3%DextranT500、KCl、磷酸缓冲液(pH7.8)和蔗糖构成的两相系统提取膜微囊的H+-ATPase活性分别被Na3VO4、KNO3、NaN3抑制了约75%、2.6%和1.3%。方向性检测显示原位膜微囊占提取质膜微囊的90%,翻转膜微囊仅占10%。去垢剂对质膜H+-ATPase活性的影响说明0.015%的TritonX-100和0.01%～0.1%的Brij58适用于测定质膜H+-ATPase活性。Lineweaver-Burk动力学分析该酶的Km值为0.65mmol·L-1,Vmax为37.59μmolPi·mg-1protein·h-1。研究结果表明:两相法提取的质膜微囊主要是正向密闭的膜微囊;胡杨愈伤组织质膜H+-ATPase的最适pH为6.5,最适温度为37℃左右。     

Suspension Culture of Endosperm Callus Cell and Amitosis In Vitro

The endosperm calli were induced on MS basic medium supplemented with lppm 2,4-D, 0.5ppm KT and 5% sucrose. The medium which contained lppm BAP, 0.1ppm NAA and 2% sucrose was used for cell suspension culture. In suspension cell culture, amitosis of cleavage division of nucleus have been observed after 5 days of culture. First the nuclear membrane and nucleolus disappeared. The crevice appeared in the center of the nucleus, and the nucleus divided into two daughter nuclei of similar size and each with a nucleolus. The daughter nucleus resembled an eye in shape. Following the emergence of cell wall, the two new unequal cells were produced. Such amitotic division proceeded repeatedly until the callus developed and eventually plantlet regenerated.