源于小孢子培养的大麦耐盐变异体获取

以大麦品种‘花30’作为供试材料,比较了甲基磺酸乙酯（EMS）和平阳霉素处理小孢子60Co γ-射线辐照处理离体穗和干种子,对300mg·L-1NaCl胁迫培养下游离小孢子的愈伤组织产量和愈伤组织在0.3％NaCl胁迫筛选下的绿苗产量的影响。结果表明,EMS处理离体小孢子和60Co γ-射线辐照千种子的愈伤组织产量和绿苗产量明显优于平阳霉素处理小孢子和60Co γ-射线辐照离体穗。以16份源于种子辐照处理的再生植株自交一代种子为供试材料,比较了在0．3％NaCl胁迫下种子的发芽率和幼苗的成活率以及植株的分蘖数、株高和单株产量。结果表明,‘花30’发芽率为0,供试的16份耐盐变异体中,有14份材料在NaCl胁迫下的发芽率优于‘花30’,鉴定出4份耐盐性明显优于‘花30’的变异体材料。选择耐盐变异体作为供试材料,测定了变异体中Na＋／H＋逆向转运蛋白基因NHXl、NHX2和NHX3和编码甜菜碱醛脱氢酶（BADH）的两个同工酶基因曰肋,和BBD2的表达模式和表达量,结果表明变异体耐盐性的提高与这些基因的表达量存在联系。     

60Coγ射线对广藿香的辐照诱变及再生植株的RAPD分析

探讨60Coγ射线辐照对广藿香外植体离体再生的影响及再生植株的诱变效应.采用60Coγ射线辐照广藿香外植体,考察辐照对外植体离体再生的影响,并观察再生植株的形态变异;同时采用随机扩增多态性DNA（RAPD）技术分析诱变再生植株的变异情况.广藿香外植体的存活率和外植体再生芽的能力,随着辐照剂量的升高而降低;部分再生植株在形态上发生一些变化;RAPD分析表明,4个辐照剂量的外植体再生植株多态性频率分别为39.13%、36.37%、23.09%及33.33%.60Coγ射线辐射会造成外植体存活率和再生芽能力的下降,辐照外植体培养获得的再生植株,在表型性状及分子水平上出现了一定的分离,为将辐照诱变技术应用于广藿香育种提供了依据.     

60Coγ射线对高免卵黄液中霉形体杀灭作用的研究

用^60Coγ射线处理高免卵黄液,研究了不同剂量、剂量率对霉形体的灭活效果和对卵黄抗体的影响。结果表明,^60Coγ射线可以完全杀灭高免卵黄液中的霉形体,霉形体在卵黄液中的灭活剂量为6．0kGy;10．0kGy以下的辐照剂量不影响卵黄液中蛋白质含量、pH值和活性氧自由基,同时不影响抗体效价。     

籼型光敏雄性不育水稻杂种花粉植株游离小孢子培养成植株

禾本科植物游离小孢子的培养已在水稻、小麦、玉米、大麦等主要农作物上获得成功,且在大麦、玉米上成功地从未经预处理及预培养的游离小孢子培养获得了再生植株。籼稻花药培养能力远远低于粳稻,对其游离小孢子的离体培养研究甚少。本文简要报道这方面的研究结果。     

芦苇^60Coγ辐照降解副产物的分析

通过^60Coγ射线处理芦苇以及分析辐照前后芦苇中的基本成分含量、副产物含量,研究辐照预处理对芦苇木质纤维素的影响.采用国标法测定了辐照前后芦苇中的纤维素、半纤维素、木质素含量;采用气质联用（GC-MS）法对芦苇^60Coγ辐照后降解副产物进行了分析.结果表明：芦苇经^60Coγ射线辐照预处理之后,纤维素和半纤维素的降解程度较大,在2 000 KGy处理处理条件下降解率分别达到83.07％和79.86％.在辐照前和辐照后的芦苇中,一共检测出42种脂肪酸类化合物、27种芳香类化合物和8种呋喃类化合物.     

60Coγ射线对高免卵黄液中EDS-76病毒灭活的研究

用不同剂量、剂量率的^60Coγ射线对卵黄液中的EDS-76病毒进行辐照,研究了其对病毒的灭活效果和对卵黄抗体(IgY)的影响。结果表明,^60Coγ射线辐照,可以灭活高免卵黄液中的EDS-76病毒,且随辐照剂量、剂量率的增大,灭活率也增高;EDS-76病毒在卵黄液中的D10值为0．57kGy～0．60kGy;6．0kGy的辐照剂量,可以将高免卵黄液中的EDS-76病毒完全灭活,且不影响卵黄抗体效价,该卵黄抗体稳定性好、耐酸、耐热、耐蛋白酶消化。     

激光—^60Co—γ辐射诱变小麦生物学效应初步研究

研究小麦品种“绵阳21号”和“汉源小麦”及其~(60)Co—γ一代(γ_1)等四个材料,经N_2和He—Ne激光辐照的20种处理(包括对照)的辐射生物学效应。试验结果表明,激光一代(L_1)植株高度,单株分蘖数,单株有效分蘖数,穗长,小穗数,不实小穗数,退化小穗数,小穗密度,主穗粒数,主穗粒重,分蘖穗粒重,单株粒重,抽穗期,叶面积,穗下轴长,倒一节间长,倒一节间粗等17种性状,都表现出不同处理所引起的生物效应差异。同株高,单株分蘖数、单株有效穗数、主穗长度、不实小穗数,主穗粒数、主穗粒重,单株粒重等8个性关变异系数的比较看出:He—Ne激光(照射10分钟)辐照干种子,“汉源小麦—γ_1”比“汉源小麦”产生的变异大,比对照(未照射)的变异更大。同时,发现不同品种所引起的抽穗期、株高、穗长等14种性状的变异分别达显著或极显著,由剂量不同引起抽穗期、主穗粒重和不实小穗数等三种性状的变异显著。     

60Coγ射线辐照对花魔芋同工酶与品质的影响

本文开展了^60Coγ射线辐照对花魔芋酶类与球茎品质影响的研究。结果表明：辐照诱发了花魔芋同工酶谱的变异,经等电聚焦电泳分析,不同剂量处理,酯酶和多酚氧化酶同工酶酶带呈现多态性,有稳定带和可变带,带型和带数的变化从分子水平揭示了魔芋辐射敏感性,是诱变效应和农艺性状突变的分子基础;经采后球茎品质检测,辐照引起了魔芋品质变化,数据分析表明干物质含量、葡甘聚糖含量、葡甘聚糖粘度与辐照剂量均呈二次曲线关系,总生物碱含量与剂量呈线性负相关。以葡甘聚糖的量和质为评价依据,中等辐射剂量范围内的辐照处理可提高魔芋品质。     

肯氏相思~(60)Co-r辐照诱变研究

采用不同剂量（3000、5000、7000Rad）(60)Co－r辐照肯氏相思种子，选育出诱变化株（M1），以M1的种子（M2）扩种试验。观测M1和M2代的形态、物候，并测定过氧化物酶同功酶带谱、酶活性、蛋白质含量，结果表明，各处理的优株与对照都存在差异。在相同生态条件下，M1代林木生长量和民代地上部生物量分别高于对照见11.48％～21．64％和14．62％～73．98％，以5000Rad辐照剂量最优，其次为7000Rad。访变个体后代表现树干直、花果期提早和种子小。     

辐射处理对冷季型草幼穗分化的影响

以C60Or射线对一年生黑麦草(品种:冠军)、多年生黑麦草(品种:爱神特)、早熟禾(品种:歌来德)及高羊茅(品种:知音)4个禾本科草坪草干种子进行辐照,剂量分别是15000rad(处理Ⅰ)、25000rad(处理Ⅱ)、35000rad(处理Ⅲ).播种生长后,田间取样观察;实验结果表明,所用剂量对冷季型草的幼穗分化进程影响不大,相对而言高剂量对多年生黑麦草的生殖生长有一定延长作用.     

蝴蝶兰EMS离体化学诱变及再生植株RAPD检测

用EMS对离体培养的蝴蝶兰类原球茎薄切片进行化学诱变,研究了不同浓度、不同时间诱变处理对类原球茎薄切片生长、类原球茎再生及再生苗生长的影响,并对再生苗DNA进行了RAPD检测。结果表明,诱变剂EMS对类原球茎薄切片生长产生重要影响,部分薄切片褐化死亡,再生类原球茎生长受到抑制,再生苗数量减少,表现出明显的伤害作用,这种伤害作用随诱变剂浓度的加大和处理时间的延长而加重。诱变剂EMS使部分再生苗畸变,叶片皱缩、变厚。并产生2株叶形、叶色突变体。部分再生苗表现出RAPD图谱带型的多态性,暗示DNA序列发生了一定的变异。EMS适合诱变剂量为浓度0．4％处理2d～4d。     

Improved isolated microspore culture efficiency in medium with maltose and optimized growth regulator combination in japonica rice (Oryza sativa)

The influence of maltose and growth regulators on microspore culture response was investigated in japonica rice. High frequency of callus induction of isolated microspores was obtained with liquid medium containing MS salts, 100 mg l^–1 myo-inositol, 1 mg l^–1 thiamine-HCl, 500 mg l^–1 glutamine, 60 g l^–1 maltose, and several growth regulators. The effect of maltose on promoting callus formation was associated with keeping a high proportion of swollen microspores after 5 day preculture and increasing the microspore division rate on the 3rd day after culture initiation. No significant effect of maltose in place of sucrose on plantlet regeneration was seen in regeneration medium. Among the growth regulators tested, the combination of auxin 2,4-dichlorophenoxyacetic acid (1 mg l^–1), naphthaleneacetic acid (1 mg l^–1), and cytokinin (6-benzyl-aminopurine 1 mg l^–1) in the medium proved to be much better for callus formation than in the other media, and the percentage of callusing microspores of that medium reached 0.86%. Indole-3-acetic acid (0.5 mg l^–1) and kinetin (2 mg l^–1) in regeneration medium were beneficial for green plantlet differentiation. The results also showed that the frequencies of microspores initial division, callus formation and green plant regeneration varied among genotypes no matter what kind of growth regulator and sugar were used. Xiushui 117 was the best variety for callusing followed by 02428 & Taipei 309. Taipei 309 showed a good ability for green plantlet regeneration.Abbreviations 2,4-d 2,4-dichlorophenoxyacetic acid - NAA naphthaleneacetic acid - 6-BA 6-benzylaminopurine - KT kinetin - IAA indole-3 acetic acid     

大麦籽粒及花药愈伤组织的游离氨基酸含量分析

对同一基因型大麦花药愈伤组织与籽粒中的游离氨基酸含量尤其是赖氨酸含量之间的关系进行分析,结果表明,对同一基因型而言,愈伤组织及籽粒中的同一氨基酸含量高低具有相同趋势。同时着重分析了愈伤组织中游离脯氨酸的含量与绿苗分化的关系,结果表明游离脯氨酸含量高的愈伤组织,其绿苗分化率较高,说明脯氨酸对绿苗分化具有重要作用。966259的花药愈伤组织及籽粒中的游离氨基酸总含量及游离赖氨酸含量均最高。     

菊芋试管苗的初代培养及愈伤组织不定芽诱导

菊芋（Helianthus tuberosus Linn.）为菊科（Asteraceae）向日葵属（Helianthus Linn.）多年生草本植物,耐寒、耐旱、耐贫瘠、耐盐碱[1];其地下块茎富含菊糖,还可通过发酵生产乙醇,在功能性食用多糖及生物能源方面的开发潜力巨大。菊芋主要通过块茎进行无性繁殖,其种子成活率和发芽率均很低[2],严重阻碍了菊芋的杂交育种。近年来以植物组织培养为基础的一系列现代育种技术为菊芋的种质改良提供了新途径,但由于菊芋的愈伤组织难以诱导不定芽或体胚发生,导致以农杆菌转化为主的转基因育种技术的应用受到限制。     

Antimitotic and hormone effects on green double haploid plant production through anther culture of Mediterranean japonica rice

Rice double haploid (DH) plants are produced mainly through anther culture. In order to improve the anther culture protocol, microspores of two japonica rice genotypes (NRVC980385 and H28) were subjected to three growth regulator combinations and four colchicine treatments on induction medium. In addition, a post anther culture procedure using colchicine or oryzalin was tested to induce double haploid plantlets from haploid plantlets. A cold pre-treatment of microspores for 9 days at 10 °C increased callus induction 50-fold in the NRCV980385 genotype. For both genotypes, 2 mg L^?1 2,4-D and 1 mg L^?1 kinetin on colchicine-free induction medium gave the best culture responses. The culturability of both genotypes changed on colchicine-supplemented induction media. A high genotype dependency was recorded for callus induction, callus regenerating green plantlets and regeneration of green double haploid plantlets. Colchicine at 300 mg L^?1 for 48 h enhanced callus induction 100-fold in H28. Colchicine-supplemented media clearly improved green double haploid plantlet regeneration. We showed that the post-anther culture treatment of haploid plantlets at 500 mg L^?1 of colchicine permitted fertile double haploid plantlets to be generated. Finally, an enhanced medium-throughput flow cytometry protocol for rice was tested to analyse all the plantlets from anther and post anther culture.     

茶树实时荧光定量PCR分析中内参基因的选择

选择合适的内参基因是提高实时荧光定量PCR分析（qRT-PCR）准确性的先决条件。该文以茶树（Camellia sinensis）芽、叶、幼根、嫩茎、花瓣、种子和愈伤组织为材料,应用实时荧光定量PCR技术,分析了18S rRNA、GAPDH、β-actin和α-tubulin4个常用内参基因在茶树不同器官组织中的表达情况。经GeNorm和NormFinder软件分析发现,当利用荧光定量PCR分析比较茶树不同器官组织中的基因表达差异时,可选择β-actin作为校正内参基因;而比较不同成熟度的叶片和愈伤组织时,可以选择GAPDH作为校正内参基因。     

甲基磺酸乙酯对水稻花药离体培养效应的研究

本文以粳稻“105”、籼稻“华03”和“鄂早6号”为实验材料,研究了不同浓度化学诱变剂EMS处理对离体培养水稻花药中花粉细胞脱分化与再分化和培养初期花药呼吸作用的影响。结果表明,低浓度EMS能显著提高花药愈伤组织诱导率,高浓度EMS则有明显的抑制作用。EMS对培养初期花药呼吸作用的影响与花药愈伤组织诱导率之间存在明显的平行关系。     

EMS和^60Co对海甘蓝种子萌发及其M1代农艺性状的影响

用ＥＭＳ及＾６０Ｃｏ处理海甘蓝种子,对其萌发种子细胞内呼吸代谢有明显的影响。低浓度的ＥＭＳ或低剂量的＾６０Ｃｏ刺激呼吸,可提高抗坏血酸氧化酶和过氧化物酶的活性,促进种子的萌发和幼苗的生长。较高剂量的ＥＭＳ和＾６０Ｃｏ抑制呼吸,与之有关的抗坏血酸氧化酶和过氧化物酶的活性,种子发芽势和发芽率、幼苗生长高度和根系活力等随之下降,用不同剂量的ＥＭＳ和＾６０Ｃｏ处理海甘蓝种子,对Ｍ１代的产量和抗病性等有一定     

Mutagenic effectiveness and efficiency of sodium azide versus ethyl methanesulfonate in maize: induction of somatic mutations at the yg2 locus by treatment of seeds differing in metabolic state and cell population.

This study was conducted to compare the effectiveness and efficiency of sodium azide tNaN3) and ethyl methanesulfonate (EMS) for inducing somatic mutations at the yg2 locus in maize seeds of two different metabolic states and cell populations. Dormant or presoaked (72 h at 20 degrees C) seeds heterozygous for yg2 locus were treated with different concentrations of either EMS or NaN3. The cell populations with respect to the percentage of cells in G1, S, G2, and M were also determined for seeds of the two metabolic states. Dormant seeds possessed a higher percentage of cells in G1 and the presoaked seeds a higher percentage of cells in S, G2, and M. The frequency of yg2 sectors in leaves 4 and 5 increased with increasing concentration of both mutagens in both dormant and presoaked seeds. Both mutagens were more effective and efficient in the presoaked seeds. NaN3 was more effective than EMS in terms of number of sectors induced per unit of dose. However, EMS was more efficient as determined by sectors induced per unit of seedling injury and clearly had the ability to induce much higher sector frequencies (more than 10 times greater) than NaN3. The low ability of NaN3 (compared to EMS) to induce mutant sectors may be related to the cells not being treated at the optimum time during the cell cycle, but it is more likely due to its low effectiveness for inducing chromosome aberrations.