耐盐突变体小麦后代耐盐稳定性分析研究

以卫星搭载小麦种子为原始材料,利用其幼穗、幼胚诱导的愈伤组织进行耐盐突变体的筛选,对耐盐愈伤组织再生植株后代进行耐盐稳定性生理生化特性分析。结果表明：(1)耐盐系后代在土壤高盐浓度条件下,游离脯氨酸含量稳定增加,且高于对照系;(2)耐盐系再生植株后代保持较高的K^ ／Na^ 比;(3)与对照相比,种子醇溶蛋白电泳带谱中的b2,b3,b5,b7带为耐盐系所特有,b8带消失;(4)耐盐系再生植株后代可溶蛋白电泳带为26条,而对照系为23条蛋白带。其中98kD、75kD、52kD、49kD和32kD为耐盐系的特有蛋白带。而38kD和35kD蛋白带为对照系所特有。     

红豆草抗羟脯氨酸变异系的筛选及其特性研究

通过选择-NaN_3诱变处理-再选择的程序从红豆草下胚轴愈伤组织获得了抗脯氨酸类似物L-羟脯氨酸的变异细胞系。抗性变异细胞系的游离脯氨酸含量比正常细胞系高4.6倍,并且抗性变异系亦表现出了很强的耐NaCl、PEG 能力。这可能为筛选抗逆境植物提供一条有效途径。     

葡萄耐羟脯氨酸变异细胞系的筛选及其特性研究（简报）

葡萄愈伤组织经甲基磺酸乙酯（EMS）诱变处理后,用羟脯氨酸（HMP）作为选择压力,筛选出耐HYP的变异细胞系。该变异系游离脯氨酸含量比正常细胞系高4．7倍,并且表现出很强的耐NaCl和PEG能力。在盐胁迫下,变异系吸收的K 和Na 含量均高于正常系,而K ／Na 比值仍与正常系相近。     

利用组织培养选择烟草耐盐愈伤组织变异体并分化出再生植株

烟草(品种革新一号)叶片为外植体,直接置入含0.5%NaCl的修改MS培养基中,诱发产生耐盐的愈伤组织。然后采取逐步提高NaCl浓度的措施,分别获得耐0.5%、1.0%、1.5%及2.0%NaCl细胞系。耐盐细胞系在无盐条件下,生长9—11代后仍保持其耐盐性。从各个耐盐细胞系均分别获得再生苗。耐2.0%NaCl的04—9细胞系共得到15株再生植株,叶片狭长、多锯齿、并具有较多的花茎,多数花粉粒畸形,经过人工授粉获得少量种子。04-9变异型再生植株水培于含有1.0—2.0%NaCl的Hogland溶液中生长85天,仍然存活。原始型愈伤组织的细胞呈不规则椭圆形,耐盐细胞系的细胞均近似圆形;耐盐浓度愈高则细胞愈小。耐盐细胞系愈伤组织的叶绿素含量随耐盐浓度增高而增加;渗透势则随耐盐水平提高而降低。耐2.0%NaCl细胞系04—9愈伤组织内脯氨酸含量高40.7倍,其再生植株叶片内的脯氨酸含量亦较原始型增加两倍。耐2.0%NaCl细胞系再生植株的幼年与成年叶片的过氧化物同工酶的酶谱与原始型均有显著差别。以上试验结果均说明耐2.0%NaCl细胞系04—9及其再生植株是一个耐盐变异体。     

耐盐药蒲公英(Taraxacum officinale Weber)愈伤组织筛选及生理生化特性分析

为获得耐1.5% NaCl的药蒲公英(Taraxacum officinale Weber)愈伤组织, 以药蒲公英叶片外植体为材料诱导愈伤组织。以NaCl为选择因子, 从愈伤组织直接筛选。在选择培养基上, 大部分愈伤组织褐化死亡, 个别褐化死亡的愈伤组织周围有少量新的细胞团长出, 将其转接到新鲜的选择培养基上, 每3周继代一次, 经3个月继代筛选获得了耐1.5% NaCl的药蒲公英细胞团。以普通愈伤组织为对照, 发现随着NaCl浓度升高, 耐盐愈伤组织的相对生长率下降但显著高于对照; 且随着盐胁迫处理时间延长持续升高, 而普通愈伤组织对照几乎停止生长, 说明耐盐愈伤组织具有相对稳定的耐盐性。在蛋白水平上, 耐盐愈伤组织与对照愈伤组织差异明显, SDS-PAGE分析显示: 耐盐愈伤组织比对照多出一条34 kD大小的蛋白带, 且30 kD、18 kD左右的蛋白带明显上调。相同处理条件下耐盐愈伤组织脯氨酸的增加幅度高于对照。盐胁迫条件下, 耐盐愈伤组织的超氧化物歧化酶(Super oxidase dimutase, SOD)、过氧化物酶(Peroxidase, POD)和过氧化氢酶(Catalase, CAT)活性明显高于对照,且随着处理时间的延长和盐浓度的增加呈现升高的趋势, 而对照则呈现先升高后下降的趋势。结果说明耐盐愈伤组织一方面通过小分子有机溶质如脯氨酸的方式调节其渗透平衡, 另一方面还可通过提高抗氧化能力降低盐分造成的次级伤害。积累蛋白也可能是耐盐愈伤组织调节渗透平衡的一种方式。通过生理生化分析确定我们获得的耐盐愈伤组织为耐盐变异体。     

枸杞耐盐突变体的筛选及生理生化分析

用不同剂量60Coγ对以枸杞叶为外植体诱导出的愈伤组织进行辐射处理,并将恢复增殖的愈伤组织采用逐步提高盐浓度的方法,直到筛选出耐1%的愈伤组织变异体,通过对其生理生化的分析表明:变异体在不同盐浓度胁迫下干、鲜重增长均高于对照组;MDA含量和质膜透性、O-·2与H2O2含量都低于对照组;抗氧化酶SOD、POD、CAT活性均高于对照组;对可溶性蛋白SDS电泳结果表明:耐盐变异体有22条蛋白带,而对照组有21条蛋白带,其中31.3kD、21.0kD、18.5kD带为耐盐变异体所特有,而74.1kD、52.1kD带为对照组所特有,并且73.7kD、67.9kD的蛋白耐盐变异体含量高于对照组.     

枸杞耐盐变异体的筛选及植株再生

枸杞(Lycium barbarum L.)无菌苗下胚轴于MS+2,4-D 0.25mg/L+LH 50 0mg/L的诱导培养基上产生胚性愈伤组织。经0.34%的EMS(半致死剂量)处理并恢复增殖2周后, 将存活组织转接到含有1.5%NaCl的诱导培养基上培养4周,再将少数存活的组织转移到含1.0%NaCl的同样培养基上继续培养,经不断选择,选出了耐1.0%NaCl的愈伤组织变异体。经耐盐性、耐盐稳定性、脯氨酸含量、叶绿素含量分析,以及对山梨醇、聚乙二醇的反应证明,该愈伤组织是耐盐变异体。变异体在含有1.0%NaCl的分化培养基(MS+6-BA0.5mg/L)上可分化出再生植株。     

诱发小麦成熟胚愈伤组织及其再生植株抗盐性变异的研究

本研究以普通小麦成熟胚为起始材料,以平阳霉素(PYM)和正定霉素(ZDM)为诱变剂。发现不同基因型对药物的敏感性不同,其中对药物敏感的在含盐筛选培养基上的存活率高。在诱导培养基内添加一定量的诱变剂可以产生耐盐变异。这种抗性愈伤组织转入与筛选培养基含盐量相同的分化培养基,比较容易产生耐盐再生植株。其M_1、M_2代较亲本系表现株高降低,穗长变短、籽粒饱满度也差;M_1代的结实率仅有5.5%,M_2代恢复到40.9%。利用M_1代和亲本系实生苗叶片,进行脯氨酸含量测定。发现耐盐变异株在无盐的Hoagland溶液中,游离脯氨酸的含量超过亲本系,对盐胁迫不敏感,其耐受力强于对照,具有一定的抗盐稳定性。     

小麦耐盐变异体的筛选Ⅱ．小麦耐盐细胞系的筛选及生理生化特性的分析

将诱导出的胚性愈伤组织采用：①逐级提高盐浓度进行胁迫筛选;②直接转入高浓度胁迫筛选;③先用羟脯氨酸（Ｈｙｐ）筛选后再转入高浓度盐中进行胁迫筛选,结果表明三种筛选方法其效果为③＞②＞①,即先用Ｈｙｐ筛选再进一步盐胁迫筛选效果最好。并对选出的耐盐细胞进行了生长量测定和生理生化特性的研究,表明耐盐细胞系脯氨酸含量不仅稳定,而且远高于对照,Ｋ￣＋／Ｎａ￣＋也大于对照,ＳＯＤ活性增加,小分子ＳＯＤ含量增加,本文并对有关问题进行了讨论。     

非洲菊耐羟脯氨酸变异系离体筛选体系的建立

以金黄色非洲菊品种深圳5号(S5)为材料,探讨了利用羟脯氨酸(Hyp)筛选耐热变异系的方法,初步建立了非洲菊耐Hyp变异系的离体筛选体系.结果表明,非洲菊新鲜花托和芽适宜的Hyp初始筛选剂量为0.4mg·mL-1,而预培养25d花托的初始筛选浓度宜增加到0.6mg·mL-1;获得的存活芽恢复培养后进行后续筛选时可以选择0.5~0.6mg·mL-1Hyp;通过对花托和芽进行5次间隔筛选后获得耐Hyp变异系SHP5,得率为0.7%.与对照(S5)相比,SHP5在室内高温胁迫条件下具有较高的游离脯氨酸含量,在自然高温条件下长势好,花序梗长度和花色素苷含量均显著增加,表现出较强的耐热性.     

卫星搭载亚麻后代中PEG和NaCl抗性系的初步筛选

把空间生物学和细胞工程相结合,通过组织培养技术对其离体筛选,得到抗１．２％ＮａＣｌ和３５％ＰＥＧ的愈伤组织,将所得抗性系愈伤组织在２．０ｍｇ／Ｌ６－苄基氨基嘌呤、０．５ｍｇ／Ｌ吲哚乙酸的ＭＳ培养基上分化得到完整的植株。抗性系能在胁迫条件下保持高的生长速度和高效的脯氨酸合成能力,表明空间诱变与组织培养相结合有望可成为筛选抗胁迫变异系的有效途径。     

卫星搭载亚麻后代中PEG和NaCl

把空间生物学和细胞工程相结合,通过组织培养技术对其离体筛选,得到抗1.2% NaCl和35% PEG的愈伤组织。将所得抗性系愈伤组织在2.0 mg/L 6-苄基氨基嘌呤、0.5 mg/L吲哚乙酸的MS培养基上分化得到完整的植株。抗性系能在胁迫条件下保持高的生长速度和高效的脯氨酸合成能力。表明空间诱变与组织培养相结合有望可成为筛选抗胁迫变异系的有效途径。     

红豆草耐盐愈伤组织的筛选及植株再生

将红豆草种子在含１．２％ＮａＣｌ的ＭＳ培养基上萌发以消除盐敏感的幼苗,把存活的幼苗下胚轴切段在含１ｍｇ／Ｌ２,４－Ｄ、０．５ｍｇ／Ｌ６－ＢＡ及１．２％ＮａＣｌ的ＭＳ培养基上诱导愈伤组织,通过连续筛选得到可耐受１．８％ＮａＣｌ的愈伤组织,在有０．２ｍｇ／Ｌ ＮＡＡ和１ｍｇ／Ｌ ＩＡＡ存在下该愈伤组织分化出芽,待幼,待幼苗长至３ｃｍ左右时转至含２ｍｇ／ＬＮＡＡ和或ＩＢＡ的１／２ＭＳ培养基上生根。对对照     

芦苇耐盐变异体与野生型植株某些特性的比较

地通过细胞工程方法得到的耐盐芦苇（Ｐｈｒａｇｍｉｔｅｓ ｃｏｍｍｕｎｉｓ Ｔｒｉｎ．）变异系Ｒ５００２－１２与野生型植株进行了分子生物学和生理生化特性比较分析。筛选到５个ＲＡＰＤ引物对变异体和野生型植株的ＤＮＡＦ的随机扩增表现出不同的多态性,表明该变异体在分子水平上发生了变化。生理生化分析的结果表明,平行生长状态下的两种株系,其可溶性蛋白和同工酶的表达水平和种类不一样,变异体在ＮａＣｌ胁迫下,表达     

Comparison of Some Characteristics Between Phragmites communis and Its Salt Tolerant Variant

R5002-12, a salt tolerant line of Phragmites communis Trin., which was obtained from ethyl methane sulfonate (EMS) treated callus selected under saline stress, was compared with its wild line in respect to their molecular biological, physiological and biochemical characterizations. Five arbitrary primers were screened which showed differences in DNA amplified polymorphism between the variant and its wild line. Some new proteins appeared in the salt tolerant plant under salt stress. Electrophoresis of peroxidase and esterase also showed some differences in isozyme expression between them. The chlorophyll content of the variant was higher than that of the original variety, whether the plants were under salt stress or not.     

The effect of NaCl on proline accumulation in potato seedlings and calli

The effects of salt stress were studied on the accumulation and metabolism of proline and its correlation with Na⁺ and K⁺ content in shoots and callus tissue of four potato cultivars, viz., Agria, Kennebec (relatively salt tolerant), Diamant and Ajax (relatively salt sensitive). Na⁺ and proline contents increased in all cultivars under salt stress. However, K⁺ and protein contents decreased in response to NaCl treatments. The activities of enzymes involved in proline metabolism, Δ¹-pyrroline-5-carboxylate synthetase (P5CS) and proline dehydrogenase (ProDH) increased and decreased, respectively, in response to elevated NaCl concentrations. The changes of P5CS and ProDH activities in more salt sensitive cultivars (Diamant, Ajax) were more than those in the tolerant ones. Then the stimulation of synthesis in combination with a partially increase of protein proteolysis, a decrease in proline utilization and inhibition of oxidation resulted in high proline contents in seedlings and calli under salt stress. In callus tissue, reduced growth and cell size may be partially responsible for high proline accumulation in response to high NaCl levels. However, although the basic proline contents in the seedlings of more salt tolerant cultivars were higher than the sensitive ones, a clear relationship was not generally observed between accumulation of proline and salt tolerance in potato.     

Production of sodium-chloride-tolerant Brassica juncea plants by in vitro selection at the somatic embryo level

Summary Somatic embryos, developed from hypocotyl segments of light-grown seedlings of Brassica juncea cv RLM198, were subjected to selection at varying concentrations of sodium chloride (NaCl). Plants were developed from proliferated somatic embryos selected on NaCl-containing medium. The selections were characterized for salt tolerance, esterase isozyme pattern, and proline accumulation. It has been found that: (i) selected tolerant lines showed better root growth, shoot growth, and fresh weight accumulation on salt-containing medium when compared to the control; (ii) salt tolerance was transmitted to the next generation in seed progeny of tolerant plants grown in the absence of exposure to salt; (iii) both the starting material and the tolerant selections accumulated proline, even when grown in salt-free medium. On salt-containing medium, however, the differences in accumulated proline between the control and tolerant lines became more pronounced, and (iv) the patterns of esterase isozymes of two tolerant selections were similar but distinctly different from that of the parental control.