兰属种间杂交胚拯救研究初报

取兰属大花蕙兰为母本、墨兰为父本进行种间杂交并进行杂种胚拯救研究。结果显示,子房离体培养明显优于胚珠离体培养,是胚抢救培养的有效手段。不同取材时间及在不同培养基中子房培养情况差异明显。在授粉后60 d子房离体培养即可获得成功,但以授粉后90 d子房离体培养最佳,萌发率高,达80%。子房培养的最适培养基为VW+BA 0.2 mg/L+NAA 0.2 mg/L+蛋白胨3 g/L+水解酪蛋白500 mg/L+椰子汁10%。     

南瓜未授粉子房离体培养及植株再生

以南瓜(Cucurbita moschata)未授粉子房为外植体, 在离体条件下研究了激素种类、基因型、胚囊发育时期、消毒方式和热激处理时间对胚状体诱导效果的影响, 旨在建立完善的南瓜未授粉子房离体诱导再生体系。结果表明, MS+4.0 mg·L^-1 2,4-D+0.5 mg·L^-1 NAA+1.0 mg·L^-1 6-BA和MS+0.04 mg·L^-1 TDZ两种培养基胚状体的诱导率都较高, 分别为19.8%和20.1%, 二者相比, 使用TDZ诱导更加简单; 在6个南瓜品种中, 2个生长势较强的品种(雪峰蜜本和鱷鱼南瓜)胚状体诱导率较高, 适合作为离体雌核诱导的实验材料; 开花当天的子房先切片后消毒可以有效降低愈伤组织的形成, 提高胚状体的诱导率; 在黑暗且35°C条件下热激5天有利于子房的转绿和出胚。将子叶胚转移至无激素的MS培养基上得到再生植株并移栽成活。经鉴定有7个再生植株的染色体数目n=x=22, 气孔保卫细胞叶绿体数的平均值为4.28个, 是单倍体植株。     

枸杞花药愈伤组织悬浮培养条件下胚状体发生与植株再生

采用枸杞花药进行离体培养,建立细胞系,诱导植株再生,结果在含不同激素的4种培养基上都诱导出了愈伤组织,诱导率为17%～169%。愈伤组织在MS 2,4D05mg/L的固体培养基上,经2～3次培养后,获颗粒状胚性愈伤组织,颗粒状胚性愈伤组织转入含相同成分的液体培养基中进行振荡培养,24h后获得大量单细胞。单细胞液经过多次继代培养,建立起稳定的悬浮系。悬浮细胞在液体培养基中培养8～10d可获得含有大量胚状体的愈伤组织块,收集悬浮培养物转移到MS 6BA02mg/L的固体培养基上,胚状体能够萌发形成大量绿色小芽,小芽转入生根培养基(MS NAA02mg/L)中20d后得到完整植株。植株根尖细胞经细胞学鉴定为单倍体。     

新疆天山雪莲体胚诱导与分化研究

以新疆天山雪莲的叶片为外植体,分别用不同配方培养基诱导愈伤组织,后进行体胚诱导和分化培养形成再生雪莲植株.结果表明,诱导愈伤组织的最适培养基为MS 2,4-D 0.5 mg/L BA 1.5 mg/L,诱导率可达到100%;愈伤组织转移至MS 2,4-D 0.5 mg/L BA 1.5 mg/L培养基进行继代培养,增殖后的愈伤组织转移到MS 2,4-D 0.2 mg/L的液体培养基后成功诱导出雪莲体胚,出胚率达40%;将体胚接至MS ABA 0.5 mg/L培养基后,结果分化生长出大量的再生雪莲幼苗.     

花生体胚诱导再生体系及基因枪转化条件的初步探讨

以花生上胚轴为外植体,研究不同Picloram(毒莠定)浓度处理和不同基因型对体细胞胚胎发生及植株再生的影响,并利用基因枪将含GUS基因的pCAMBIA2301质粒载体轰击体胚,对基因枪转化条件进行了初步探索.结果表明:外植体在添加5 mg/ L Picloram+1 mg/L Glutamine(谷氨酰胺)的MB培养基上诱导的体胚发生率、产胚数及植株再生率最高.不同基因型以‘中花8号'体胚再生率最高(体胚诱导率为55.36%,植株再生率为56.79%).在氦气压力为1 100 psi,轰击距离为9 cm时,体细胞胚GUS瞬时表达率可达到22.95%.     

江西铅山红芽芋胚性愈伤组织的包埋玻璃化超低温保存

为长期安全保存江西铅山红芽芋种质资源,本文以江西铅山红芽芋的胚性愈伤组织为对象,研究了包埋玻璃化冻存过程中各因素对细胞活力和愈伤组织成活率的影响,优化建立了江西铅山红芽芋胚性愈伤组织包埋玻璃化超低温保存体系。将约0.2 g胚性愈伤组织块包埋成海藻酸钙凝胶珠后,在25℃下转入MS+2 mg/L TDZ+1 mg/L NAA+0.75 mol/L蔗糖的培养基中于14 h/d光周期下预培养1 d;预培养后的胚性愈伤组织块用2 mol/L甘油和0.4 mol/L蔗糖的混合物在25℃下装载40 min;采用PVS2在25℃下脱水30 min,更换PVS2后直接投入液氮保存1 d;再将胚性愈伤组织块置于37℃恒温水浴中化冻3 min,然后用MS+2 mg/L TDZ+1 mg/L NAA+1.2 mol/L蔗糖的液体培养基洗涤3次,每次10 min;洗涤后的胚性愈伤组块转入MS+2 mg/L TDZ+1mg/L NAA固体培养基上先暗培养7 d再转到14 h/d光周期中培养。7 d后胚性愈伤组织块开始恢复生长,并且在30 d内分化出胚状体;将胚状体再次转入MS+2 mg/L TDZ+1 mg/L NAA固体培养基上,60 d后形成完整的植株。红芽芋胚性愈伤组织包埋玻璃化超低温保存后的平均成活率约为60%,并且红芽芋胚性愈伤组织冻后再生苗没有发生形态性状和染色体数目的变异,此结果为长期安全保存江西铅山红芽芋种质资源奠定了良好的基础。     

白皮松成熟胚的离体培养研究

以白皮松成熟胚为外植体诱导再生小植株.试验结果表明,成熟胚不定芽诱导以MS培养基最佳,附加0.294 mg/L的NAA和3.56 mg/L的6-BA时,诱导率接近100%;MS培养基附加NAA(0.05 mg/L)时,平均增殖系数可达6.3以上.不定芽增殖率最大值(10)出现在SH培养基上,此时NAA浓度为0.05 mg/L、6-BA浓度为4 mg/L.MS培养基中加入适量活性炭和GA3能促进不定芽生长,随着活性炭和GA3浓度的增加,有效嫩梢(≥2cm和≥4 cm)的比率显著增高;当活性炭和GA3浓度过高时(分别超过2.75 g/L和4.1 mg/L),不定芽的伸长与生长受到抑制.在离体培养条件下,以种胚为外植体获得了无根苗.     

酿酒葡萄"梅尔诺"再生系统建立的研究

以酿酒葡萄“梅尔诺”离体胚珠、叶柄为材料．通过控制激素水平、光照和温度等,对建立再生体系的器官发生途径和体胚发生途径进行了研究。结果表明,体胚的诱导和不定芽的再生与基本培养基、叶柄的着生部位、生长调节物质种类和浓度等因素有关。由“梅尔诺”的胚珠愈伤组织再生出体细胞胚的最佳培养基配方为CPSE培养基(CP287 BA 0．2mg／L NOA 1．0mg／L),体细胞胚再生率可达47．50％。“梅尔诺”体细胞胚在CPSE培养基上100％萌发为芽状,将其切断置于培养基MS TDZ 4．0mg／L上可直接诱导出绿色不定芽,再生率为52．25％;同时在培养基MS TDZ2．0mg／L上获得了“梅尔诺”离体叶柄再生不定芽．再生率为62．42％．二者再生的不定芽的最他增殖培养基为MS BA0．5mg／L。“梅尔诺”体细胞胚的萌发芽在WPM培养基中能很好的生根及成苗,并建立了单芽茎段微繁体系。     

小麦成熟胚愈伤组织诱导及分化研究

以2个小麦品种成熟胚为外植体进行离体培养,研究了不同预处理、不同2,4-D浓度及与KT组合、不同蔗糖浓度等因素对愈伤组织诱导及分化的影响。结果表明:4℃低温预处理可提高愈伤组织的出愈率及再生苗率,2个材料的出愈率及再生苗率均达到90%和30%以上;在不同预处理条件下,2,4-D浓度对出愈率及再生苗率的影响与基因型有关,2,4-D浓度为1~2 mg/L更有利于愈伤组织诱导及分化;附加KT能缓解高浓度2,4-D对再生苗率的抑制作用,而对于在1、2 mg/L 2,4-D的培养基中附加KT则不表现这种作用;蔗糖浓度则在30 g/L条件下更有利于愈伤组织诱导。因此通过4℃低温预处理,在MS基本培养基中附加1~2mg/L 2,4-D及30 g/L蔗糖亦可促进小麦成熟胚愈伤组织的诱导和分化。     

‘光叶蔷薇’器官间接再生体系的建立及GUS基因转化的初步研究

以‘光叶蔷薇’(Rosa wichuriana‘Basye's thornless’)无菌苗的顶生幼嫩小叶为外植体,探讨了其愈伤组织诱导及植株再生的方法。结果表明,高浓度的生长素NAA能诱导外植体产生愈伤组织;由NAA诱导的愈伤组织在附加TDZ的MS培养基上,先暗培养再进行光照培养可直接分化出不定芽。诱导愈伤组织的最佳NAA浓度是7.0 mg/L、暗培养时间为10 d,而最佳分化培养基是MS+5.0 mg/L TDZ+30 g/L葡萄糖+2.5 g/L GEL,分化率达18.34%。以诱导产生的愈伤组织为侵染受体,初步建立了‘光叶蔷薇’GUS基因转化体系。农杆菌菌液浓度OD600值为0.5、侵染30 min、共培养2 d、乙酰丁香酮的浓度为50μmol/L是‘光叶蔷薇’愈伤组织转基因的最优条件。     

Investigation of the mechanism of temperature sensitivity of the electroreceptors of ampullae of lorenzini

In experiments on Black Sea skates (Raja clavata), the potential of the receptor epithelium of the ampullae of Lorenzini and spike activity of single nerve fibers connected to them were investigated during electrical and temperature stimulation. Usually the potential within the canal was between 0 and –2 mV, and the input resistance of the ampulla 250–400 k. Heating of the region of the receptor epithelium was accompanied by a negative wave of potential, an increase in input resistance, and inhibition of spike activity. With worsening of the animal's condition the transepithelial potential became positive (up to +10 mV) but the input resistance of the ampulla during stimulation with a positive current was nonlinear in some cases: a regenerative spike of positive polarity appeared in the channel. During heating, the spike response was sometimes reversed in sign. It is suggested that fluctuations of the transepithelial potential and spike responses to temperature stimulation reflect changes in the potential difference on the basal membrane of the receptor cells, which is described by a relationship of the Nernst's or Goldman's equation type.I. P. Pavlov Institute of Physiology, Academy of Sciences of the USSR, Leningrad. I. M. Sechenov, Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Pacific Institute of Oceanology, Far Eastern Scientific Center, Academy of Sciences of the USSR, Vladivostok. Translated from Neirofiziologiya, Vol. 12, No. 1, pp. 67–74, January–February, 1980.     

Principles of organization and evolution of systems of regulation of functions

Evolution of living organisms is closely connected with evolution of structure of the system of regulations and its mechanisms. The functional ground of regulations is chemical signalization. As early as in unicellular organisms there is a set of signal mechanisms providing their life activity and orientation in space and time. Subsequent evolution of ways of chemical signalization followed the way of development of delivery pathways of chemical signal and development of mechanisms of its regulation. The mechanism of chemical regulation of the signal interaction is discussed by the example of the specialized system of transduction of signal from neuron to neuron, of effect of hormone on the epithelial cell and modulation of this effect. These mechanisms are considered as the most important ways of the fine and precise adaptation of chemical signalization underlying functioning of physiological systems and organs of the living organism