水稻两性生殖细胞的N-甲基-N-亚硝基脲诱变方法

N-甲基-N-亚硝基脲(MNU)被用于水稻(Oryza sativa)受精卵的诱变。通过水稻辽盐6号成熟生殖器官的MNU体内同步处理及后代群体筛查, 确立了水稻两性生殖细胞的MNU诱变方法。与辽盐6号受精卵的MNU处理相比, 各组条件下两性生殖细胞的MNU处理明显使M₁群体生长发育的指标降低及M₁-M₂群体中突变性状的发生率升高。两性生殖细胞在含有1.5 mmol∙L ^-1 MNU和10 mmol∙L ^-1 PO₄ ^3-的缓冲液(pH4.8)中处理60分钟, 突变性状发生率是基于受精卵MNU处理的3倍。进一步筛查M₃群体, 获得了包含新型植株和籽粒突变体的纯合突变体系列。研究结果表明, 水稻两性生殖细胞的MNU诱变可显著提高广谱诱变效率。该技术的应用可为水稻的未知功能基因鉴定和育种所需的各种突变体规模化开发提供高效的技术支撑。     

一个新的水稻叶绿素缺失黄叶突变体的特征及基因分子定位

从粳稻“嘉花1号”⁶⁰Coγ射线辐照的后代中筛选到一个叶绿素缺失黄叶突变体(yl11), 与野生型“嘉花1号”相比该突变体表现为全生育期植株叶片呈黄色, 叶绿素含量以及净光合速率明显下降, 叶绿体发育不完善, 并且伴随着株高等主要农艺性状的变化。遗传分析表明, 该突变性状受一对隐性核基因(yl11)控制。该突变体与籼稻“培矮64S”杂交生产的F₂、F₃群体中的分离出突变体型920个单株作为定位群体, 利用SSR和InDel分子标记将yl11基因定位在水稻第11染色体长臂上的MM2199和ID21039分子标记之间, 其物理距离约为110 kb, 目前该区域内没有发现与水稻叶绿素合成/叶绿体发育相关已知功能基因。研究结果为今后对该基因的克隆和功能分析奠定了基础。     

植物生长调节剂对银柴胡细胞生长特性的影响

利用细胞悬浮培养技术,研究不同激素对银柴胡细胞生长特性及过氧化氢酶和硝酸还原酶活性的影响。结果表明,第4种激素配比的培养液M₄(MS+6-BA 1.0 mg·L^-1+2,4-D 1.0 mg·L^-1+NAA 1.0 mg·L^-1)的细胞鲜重、干重,细胞数目均明显高于其他处理;M₄处理可有效增强银柴胡细胞硝酸还原酶的活性,提高氮素利用率;M₂(MS+6-BA 1.0mg·L^-1+NAA0.5 mg·L^-1)处理有利于增强银柴胡细胞过氧化氢酶活性。     

基于FvCB模型分析盐分胁迫对棉花叶片光合作用的影响

为深入理解叶片光合特性对盐胁迫的响应机理,以棉花为试验材料,设置5个盐分(NaCl)浓度处理:0(CK)、50、100、150和200 mmol·L^-1,利用FvCB模型分析盐胁迫对棉花幼苗叶片光合特性的影响。结果表明:与CK相比,50和100 mmol·L^-1盐分处理增加了棉花叶片的最大羧化速率(V_{c max})和最大电子传递速率(J_max),但150和200 mmol·L^-1盐分处理显著降低了V_{c max}和J_max。叶片净光合速率(P_n)、叶肉导度(g_m)和暗呼吸速率(R_d)随盐分浓度升高而下降;与CK相比,50和100 mmol·L^-1盐分处理对g_m无显著影响,但P_n和R_d显著降低。150和200 mmol·L^-1盐分处理明显降低了P_n、g_m和R_d,且与0、50和100 mmol·L^-1盐分处理间存在显著差异;利用FvCB模型模拟了不同盐分胁迫下叶片净光合速率。与不考虑g_m的模拟结果相比,考虑g_m提高模拟值和实测值间的决定系数,并降低了平均绝对误差。棉花幼苗耐盐阈值为100～150 mmol·L^-1,随盐分浓度的增加,光合限制因素由叶肉因素转变为光合机构受损;引入g_m可以提高FvCB模型的模拟精度。     

盐胁迫下外源Ca2+对花生生长发育、生理及产量的影响

以‘花育22’为试验材料,使用外源钙[0、6、12 mmol·L^-1的Ca(NO₃)₂]处理盐胁迫(100 mmol·L^-1 NaCl)及正常条件下生长的花生,以盆栽方式研究了不同Ca²⁺浓度处理对盐胁迫条件下花生整个生育期的相关生理与产量指标的影响.结果表明: 在100 mmol·L^-1 NaCl条件下,施加不同浓度外源钙均可提高超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性以及叶绿素含量,降低丙二醛(MDA)含量和电解质外渗,增加根系活力,改善植株的农艺性状,增加生物积累量,最终提高花生产量,并且12 mmol·L^-1 Ca²⁺处理的效果最显著.通过增强活性氧的清除能力、维持细胞膜的稳定性以及完整性,是外源钙有效缓解花生植株的盐胁迫伤害并最终提高荚果产量的重要原因.     

盐胁迫下外源Ca2+对花生生长发育、生理及产量的影响

以‘花育22’为试验材料,使用外源钙[0、6、12 mmol·L^-1的Ca(NO₃)₂]处理盐胁迫(100 mmol·L^-1 NaCl)及正常条件下生长的花生,以盆栽方式研究了不同Ca²⁺浓度处理对盐胁迫条件下花生整个生育期的相关生理与产量指标的影响.结果表明: 在100 mmol·L^-1 NaCl条件下,施加不同浓度外源钙均可提高超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性以及叶绿素含量,降低丙二醛(MDA)含量和电解质外渗,增加根系活力,改善植株的农艺性状,增加生物积累量,最终提高花生产量,并且12 mmol·L^-1 Ca²⁺处理的效果最显著.通过增强活性氧的清除能力、维持细胞膜的稳定性以及完整性,是外源钙有效缓解花生植株的盐胁迫伤害并最终提高荚果产量的重要原因.     

有机肥氮投入比例对双季稻田根际土壤微生物生物量碳、氮和微生物熵的影响

为探明不同有机肥氮素占总氮投入的百分比对双季稻区早、晚稻各生育时期稻田根际土壤微生物的影响,本研究以大田定位试验为平台,应用氯仿熏蒸-K₂SO₄提取法和化学分析法系统分析了施用化肥N(M₁)、30%有机肥N(M₂)、50%有机肥N(M₃)、100%有机肥N(M₄)和无N对照(M₀)5个不同施肥处理双季稻田根际土壤微生物生物量碳(MBC)、微生物生物量氮(MBN)和微生物熵的差异.结果表明: 在早稻和晚稻各主要生育时期,施肥措施均能提高稻田根际土壤MBC、MBN和微生物熵,各施肥处理根际土壤MBC、MBN和微生物熵均随水稻生育期推进呈先增加后降低的变化趋势,均于齐穗期达到最大值,成熟期为最低值;其中,各处理双季稻田根际土壤MBC、MBN、MBC/MBN值和微生物熵一般均表现为M₄＞M₃＞M₂＞M₁＞M₀,M₂、M₃和M₄处理间均无显著差异,但均显著高于M₀处理.可见,单独施用化肥措施对提高根际土壤微生物生物量碳、氮和微生物熵效果有限,施用有机肥或有机无机肥配施提高根际土壤微生物生物量碳、氮和微生物熵的效果较好.     

激光对家蚕诱变效应的探讨

激光应用于生物学的研究,国内外都取得 了一定进展。Zkzolot^[8]用红宝石激光器照射雄 性果蝇幼虫产生了突变。中山大学^[3]用CO₂激 光器处理植物花药,引起大量染色体畸变。安 徽农学院^[5]用钦玻璃激光器照射蚕卵,获得一 雌一雄的突变体,进而育成新原种。华南农学 院^[2]用显离子激光照射蓖麻蚕蛹,诱发蛾翅突 变,并育成优良新品种。但也有经照射后未发 现突变的>,对此有必要从实践和理论上作进 一步探讨。我们采用特定位点法,以家蚕形质 性状为标志,研究了激光对家蚕诱变的效应,并 进行了染色体观察,现将结果报告如下.     

Construction of CO2-response model of electron transport rate in C4 crop and its application

准确估算光合电子流对CO₂响应的变化趋势对深入了解光合过程具有重要意义。该研究在植物光合作用对CO₂响应新模型(模型I)的基础上构建了电子传递速率(J)对CO₂的响应模型(模型II), 并对用LI-6400-40便携式光合仪测量的玉米(Zea mays)和千穗谷(Amaranthus hypochondriacus)的数据进行了拟合。结果表明, 模型II可以很好地拟合玉米和千穗谷叶片J对CO₂浓度的响应曲线(J-C_a曲线), 得到玉米和千穗谷的最大电子传递速率分别为262.41和393.07 mmol·m ^-2·s ^-1, 与估算值相符合。在此基础上, 对光合电子流分配到其他路径进行了探讨。结果显示, 380 mmol·mol ^-1 CO₂浓度下玉米和千穗谷碳同化所需的电子流为247.92和285.16 mmol·m ^-2·s ^-1, 分配到其他途径的光合电子流为14.49和107.91 mmol·m ^-2·s ^-1(考虑植物CO₂的回收利用)。比较两种植物的其他途径光合电子流分配值发现, 两者相差6倍之多。分析认为这与千穗谷和玉米的催化脱羧反应酶种类以及脱羧反应发生的部位不同密切相关。该发现为人们研究C₄植物中烟酰胺腺嘌呤二核苷磷酸苹果酸酶型和烟酰胺腺嘌呤二核苷酸苹果酸酶型两种亚型之间的差异提供了一个新的视角。此外, 构建的电子传递速率对CO₂的响应模型为人们研究C₄植物的光合电子流的变化规律提供了一个可供选择的数学工具。     

长白落叶松体胚发生再生体系优化

以长白落叶松(Larix olgensis)未成熟合子胚为外植体诱导胚性愈伤组织, 通过调节影响体胚发生的营养物质和植物生长调节剂配比, 进行愈伤组织的胚性恢复与保持以及体胚发生再生体系的优化。结果表明: 不同无性系之间胚性愈伤组织诱导率差异显著, 胚性愈伤组织在S+0.2 mg·L ^-1NAA+0.5 mg·L ^-1BA+0.5 mg·L ^-1KT+0.5 g·L ^-1谷氨酰胺+0.5 g·L ^-1水解酪蛋白+30 g·L ^-1蔗糖及3.0 g·L ^-1植物凝胶培养条件下, 可以恢复胚性并长久保持。在S+20 mg·L ^-1ABA+60 g·L ^-1PEG₄₀₀₀+60 g·L ^-1蔗糖及3.0 g·L ^-1植物凝胶条件下分化培养6周, 体胚发生率可达100%。将正常发育的体胚先在WPM+ 6 mg·L ^-1间苯三酚+1.0 g·L ^-1活性炭+3.0 mg·L ^-1VB₁+20 g·L ^-1蔗糖及3.0 g·L ^-1植物凝胶条件下培养2周, 再转接至B₅+ 0.4 mg·L ^-1NAA+1.0 mg·L ^-1IBA+0.5 mg·L ^-1GA₃+2.0 mg·L ^-1VB₁+1.0 g·L ^-1活性炭+20 g·L ^-1蔗糖及3.0 g·L ^-1植物凝胶条件下培养2周, 可见子叶舒展、下胚轴伸长且根系正常的体胚苗。该研究建立了长白落叶松胚性愈伤组织胚性恢复与保持方法, 并进一步优化了体胚发生的植株再生体系, 为林木资源快速繁育和遗传改良奠定了基础。     

Spring Time and Autumn Time Toxic Effects of Copper (II), 3-(2-Furyl) Prop-2-Enal Semicarbazone and [CuCl2(FASC)2] Complex in Mice

In vitro, 3-(2-furyl) prop-2-enal semicarbazone-copper (II) complex [CuCl₂(FASC)₂] presents antimitotic effects. In this work we studied the in vivo seasonal toxic effects in male Swiss mice of CuCl₂, and FASC and the [CuCl₂(FASC)₂] complex. In spring, one injection of CuCl₂8.10^-2 mmol killed 16% of animals after 24 h. Cupric chloride lethal dose was up to 64.10^-2 mmol with 100% mice dead after 24 h. FASC was well tolerated from 0.65 to 1.3 mmol. The complex was 100% lethal with 48.10^-2 mmol. In autumn, mice were more sensitive to CuCl₂ and to the complex with lethal doses up to 32.10^-2 mmol and 8.10^-2 mmol, respectively. On the other hand, FASC was well tolerated. It is concluded that the in vivo toxic effects of CuCl₂ and [CuCl₂(FASC)₂] complex are quite different in spring and autumn.     

Synthesis, NMR spectroscopy study, and antimuscarinic activity of a series of 2-(Acyloxymethyl)-1,3-dioxolanes

A series of 1,3-dioxolane-based ligands, bearing hydroxymethyl or ester functionalities, was synthesized and tested as potential muscarinic antagonists. The compounds display moderate to low affinity for the three receptor subtypes M₁-M₃, with some of them showing a significant selectivity for the M₃ subtype. The configurational and conformational properties were studied using NOE experiments and vicinal coupling constants. The ¹H and ¹³C NMR chemical shifts show stereochemically dependent trends. Quantitative analysis of conformer populations showed that the exocyclic CH₂N⁺(CH₃)₃ group is prevalently in a pseudo-axial orientation in the cis isomers and in a pseudo-equatorial orientation in the trans isomers.     

水杨酸调控盐胁迫下羽衣甘蓝种子萌发的机理

盐胁迫是植物种子萌发与植株生长的重要限制因子。以羽衣甘蓝(Brassica oleracea var.acephala)名古屋为材料,研究不同盐分对其种子萌发的影响,探索水杨酸(SA)及其合成抑制剂氨基茚磷酸(AIP)处理对羽衣甘蓝种子萌发的调控效应。实验结果表明,150与200 mmol·L^–1 NaCl处理后的羽衣甘蓝种子活力显著降低。盐胁迫显著降低种子的吸水速率、种子活力与幼苗质量,降低苯丙氨酸裂解酶活性与内源SA含量,提高过氧化氢(H2O2)与超氧阴离子(O2^–.)含量。SA可以缓解盐胁迫对羽衣甘蓝种子活力的抑制作用,通过促进内源SA合成,从而提高种子吸水率与种子活力,促进种子对K^+、Mg^2+的吸收,降低Na+含量。此外,外源施加SA能够显著增强超氧化物歧化酶和过氧化物酶活性,降低H2O2与O2^–.的积累。相反,氨基茚磷酸(AIP)处理能够增强盐胁迫对种子萌发的抑制作用,推测这与AIP处理能够显著降低种子内源SA含量密切相关。研究表明外源SA主要通过提高保护酶活性、降低活性氧积累和维持体内离子平衡来增强羽衣甘蓝的耐盐性。     

外源物质对茶树耐寒及蔗糖代谢关键基因表达的影响

冬春季节低温伤害是影响茶树(Camellia sinensis)生产的重要因素。以茶树盆栽苗为试验材料,通过喷施不同浓度的γ-氨基丁酸(GABA)、绿藻粉和竹醋液溶液,研究低温胁迫下3种外源物质对茶树耐寒能力的影响,并对蔗糖代谢关键基因SPS、SUS4、INV4和INV5表达量及耐寒相关生理指标进行分析,解析外源物质影响茶树耐寒性的生理与分子机制。结果表明,低温胁迫下,用不同浓度GABA、绿藻粉和竹醋液喷施处理茶树盆栽苗叶片,其冻害指数和相对电导率显著低于对照;可溶性糖含量显著高于对照,以10 mmol·L^–1 GABA、0.22 mg·mL^–1绿藻粉及2.5 mg·mL^–1竹醋液处理效果最佳。低温胁迫下,与清水处理相比,分别用10 mmol·L^–1 GABA、0.22 mg·mL^–1绿藻粉或2.5 mg·mL^–1竹醋液处理茶树盆栽苗后,茶树苗丙二醛含量显著降低,抗氧化酶活性显著提高;叶片叶绿素、可溶性糖和脯氨酸含量显著增加,蔗糖含量在处理72小时后分别增加了15.24%、11.39%和5.97%;SPS、SUS4、INV4和INV5基因的表达量显著升高。实验结果表明,GABA、绿藻粉和竹醋液能显著增强茶树的耐寒性。研究结果可为茶树抗寒剂的筛选提供理论依据。     

Response of Escherichia coli to ethyl methanesulfonate: Influence of growth phase and repair ability on survival and mutagenesis

The effects of altering the cell growth rate (physiological state) and DNA repair capacity (genetic state) on susceptibility to inactivation and mutagenesis by ethyl methanesulfonate (EMS) were studied in 4 strains of E. coli. Logarithmic and stationary phase cells of the polymerase I deficient mutant, P₃₄₇8 polA, a recombination deficient mutant, DZ₄₁₇ recA, and the respective parental strains, W₃₁₁₀pol⁺ and AB₂₅₃ rec⁺, were exposed to EMS and the surviving fraction and mutant frequency determined. At the same EMS concentration both mutants were more susceptible to inactivation than the parental strains. In all 4 strains, log phase cells were more sensitive to inactivation than stationary cells. The surviving fraction of stationary cells exceeded log cells by a factor of 18 for polA, 6 for recA, and about 2 for the parental strains. In all strains, except recA, log phase cells exhibited higher spontaneous mutant frequencies than stationary phase cells. At the same concentration of EMS, survivors of both polA and recA showed more than 10-fold higher induced frequencies than the wild types. However, at the same survival levels the repair deficient mutants exhibited induced mutant frequencies comparable to the repair proficient strains. There was no significant effect of growth phase on EMS induced mutability in recA or the parental strains. In marked contrast, the polymerase I deficient mutant shows both a higher spontaneous frequency and a greater than 10-fold higher EMS induced mutant frequency in log phase cultures compared to stationary phase cultures. Our results support the hypothesis that cellular susceptibility to alkylating agents is influenced by both the genetic capability for repair and the particular physiological state of the cell.     

Characterization and Separation of Some of the Coordination Compounds of Chromium and Aluminon

Chromic nitrate and aluminon (ATA) react to form highly colored lakes. Examination of these lakes by the Vosburgh and Cooper method has led to the identification of a pigment [(ATA)Cr(H₂O)_x] which is insoluble in water and absolute alcohol, a deep red anionic component [(ATA)_BCr(H₂O)_x]^-y, and a purplish-red cationic component [(ATA)Cr₄(H₂O)_x]^+y. These components of the lake have been isolated and separated in the dry form. The anionic compound is apparently a simple coordination compound and can be used as a cytoplasmic stain while the cationic component is a chelate and can be used as a rather selective nuclear stain. In addition to these components, a number of other components were also found. Not only the ratio between chromium and aluminon but also the concentration of the reactants influences the formation of these different components. The amount of pigment formed is maximal with a molar ratio of 1 and a concentration of 10^-2 M or 10^-1 M. The anionic component is maximal with a molar ratio of 1 Cr to 6 aluminon at the 10^-1 M concentration, the cationic component is maximal with a molar ratio of 4 chromium to 1 aluminon at the 10^-1 M concentration. None of these are formed at the 10^-4 M concentration but only a deep redpurple soluble compound at the 1 : 1 ratio, which was not further investigated.     

Enzymatic synthesis of new aromatic and aliphatic esters of flavonoids using Candida antarctica lipase as biocatalyst

Enzymatic acylation of rutin and esculin with aromatic, aliphatic and aryl-aliphatic acids using Candida antarctica lipase in tert amyl alcohol as solvent was investigated under low water content. Whatever the acyl donor used, the conversion yields and initial rates for esculin were higher than for rutin. For a given flavonoid, the performance of these reactions depended on the acyl donor structures. For aliphatic acids, conversion yields and initial rates of both flavonoids were respectively in the ranges of 68-90% and of 9.5×10^-2-72×10^-2 mmol l^-1 h^-1. For aromatic acids, the reaction occurred only with the aryl subgroup (cinnamic, hydrocinnamic, 3,4-dihydroxyhydrocinnamic and 4-hydroxyphenyl acetic acids) and was drastically influenced by the presence of side chain and substitution patterns of the aromatic ring. Except for hydrocinnamic acid (75%, 23.4×10^-2 mmol l^-1 h^-1), with these acids the conversion yields and initial rates were lower and in the range of 10-45% and of 0.7×10^-2 to 12.1×10^-2 mmol l^-1 h^-1. Unsaturation of the side chain of the hydrocinnamic acid decreased the esculin conversion rate from 75 to 13% and initial rate from 23.4 to 1.76×10^-2 mmol l^-1 h^-1. The presence of hydroxyl or nitro-groups on the aromatic ring of the aryl aliphatic acid also reduced conversion yields and initial rates. Even without a spacer, the non-phenolic ring acid (quinic acid) was reactive and lead to conversion yields of about 20 and 23% respectively for rutin and esculin.