龙凤竹的组织培养

以龙风竹[Pedilanthus tithymaloides（L.）Poit．var．nartus Dressler]茎段为外植体,研究了龙凤竹愈伤组织诱导、植株再生以及试管苗继代保存培养的培养条件。结果表明,龙风竹茎段灭菌的最佳方法是用1．0g·L^-1HgCl2处理4～10min;愈伤组织诱导与分化的最佳培养基为添加1．5mg·L^-1 6-BA和0．10～0．15mg·L^-1 NAA的MS培养基（含有30g·L^-1蔗糖和6g·L^-1琼脂粉,pH5．78～pH5．80）;试管苗生根的最佳培养基为含有0．2mg·L^-1 NAA的生根培养基（1／2MS,含有15g·L^-1蔗糖和6g·L^-1琼脂粉,pH5．78-pH5．80）,试管苗生根率可以达到93．3％;经过炼苗并移栽后,龙风竹试管苗的成活率可达95．0％以上;龙凤竹试管苗的最佳继代保存培养条件为：在含有0．1mg·L^-1 NAA的生根培养基中,于温度15℃、光照强度20μmol·m^-2·s^-1的条件下继代保存。此外,龙凤竹愈伤组织可以直接分化产生大量丛生芽,达到龙凤竹试管苗增殖的目的。     

AgNO3对网纹甜瓜试管苗糖代谢及抗氧化酶活性的影响

在网纹甜瓜继代培养过程中加入不同浓度的AgNO3,测定试管苗糖代谢及抗氧化酶活性等。结果表明,玻璃苗果糖、葡萄糖和可溶性糖含量显著降低,而蔗糖含量显著高于正常苗;抗氧化系统发生紊乱,O2产生速率降低,H2O2含量下降,MDA含量增加,木质素含量降低。加入30μmol·L^-1AgNO,使玻璃苗蔗糖合成酶、蔗糖转化酶活性和木质素含量与正常苗差异不显著。说明适当浓度AgNO3有利于维持试管苗糖代谢正常进行,降低试管苗脂质过氧化程度,从而降低或抑制玻璃化现象的发生。     

AgNO3逆转的黄芩组培玻璃化苗的活性成分含量及其生物活性研究

研究AgNO3对黄芩组培玻璃化苗的逆转作用,并在此基础上对逆转的黄芩组培玻璃化苗的活性成分含量及活性进行分析。结果表明,AgNO3对黄芩的玻璃化苗具有明显的逆转作用,且这种逆转作用受AgNO3浓度的影响。在AgNO3的浓度为4 mg·L^-1时,玻璃化苗的逆转率最高为88%,且逆转的黄芩苗的生长状态最好。在此基础上,对逆转1个月的黄芩玻璃化苗的活性成分含量及生物活性进行研究。逆转的黄芩玻璃化苗体内黄芩苷的含量为68.6 mg·g^-1,明显高于玻璃化苗(38.3 mg·g^-1),但低于正常的组培苗(108.2 mg·g^-1)。此外,提取物的抗氧化、抗菌和抗肿瘤实验结果显示,逆转的黄芩玻璃化组培苗明显好于玻璃化苗,但仍低于正常的组培苗。以上研究结果表明,AgNO3对黄芩组培玻璃化苗具有较好的逆转及恢复作用。     

不同浓度卡那霉素、潮霉素对楸树试管苗生长的影响

目的:将不同质量浓度的卡那霉素、潮霉素加入楸树培养基中,研究卡那霉素、潮霉素对楸树组培苗生长的影响,以确定抗生素对楸树茎段分化与生根的敏感质量浓度。方法:待楸树继代、生根培养基灭菌后温度降至30~50℃,将不同质量浓度的卡那霉素、潮霉素经抽滤式灭菌加入培养基中,在培养基中接入楸树组培无菌茎段培养,观测茎段继代(增殖芽数、芽长、叶数等)、生根(发根数、根长、芽长等)生长指标并统计分析。结果:楸树组培继代培养基添加卡那霉素质量浓度为100 mg/L时组培瓶苗生长缓慢,浓度为150 mg/L时叶片大部分发白并干枯,茎段基部无愈伤组织形成,瓶苗基本停止生长,楸树继代瓶苗对卡那霉素耐受性范围为100~150 mg/L;添加潮霉素质量浓度为5 mg/L时瓶苗生长较为缓慢,浓度为10 mg/L时叶片开始干枯,茎段基部愈伤组织较小,瓶苗基本停止生长,楸树继代瓶苗对潮霉素耐受性范围为10 mg/L左右。楸树组培生根培养基添加卡那霉素质量浓度为100 mg/L时大部分茎段干枯,少部分为绿但未分化芽与根,浓度为150 mg/L时大部分茎段干枯,极少上部为绿,基部干枯,但未分化芽与根,楸树组培瓶苗生根培养苗对卡那霉素耐受性范围为100~150 mg/L;添加潮霉素质量浓度为5 mg/L时少部分茎段干枯,浓度为10 mg/L时大部分茎段干枯,少部分为绿,茎段未出现芽的分化与根的萌发现象,楸树组培瓶苗生根培养苗对潮霉素耐受性范围为5~10 mg/L。结论:卡那霉素、潮霉素对楸树组培苗生长有明显的抑制作用且与抗生素浓度呈负相关,但低质量浓度(1 mg/L)的潮霉素对楸树继代分化芽数有促进作用;同一抗生素对楸树不同无性系间组培苗生长的影响无显著差异。     

樟叶越桔组培苗生根和移栽技术研究

为解决樟叶越桔(Vaccinium dunalianum)组培苗生根质量不佳、移栽成活率低的问题,该研究以樟叶越桔继代苗为材料,采用单因子试验从激素类型及浓度、培养基类型和蔗糖质量浓度对其生根的适宜条件进行筛选,进一步研究了不同基质配比对樟叶越桔移栽苗存活率的影响。结果表明:激素类型和浓度、培养基类型对樟叶越桔生根率的影响最大,其次为蔗糖质量浓度;最适合樟叶越桔生根的激素及浓度为IBA2.0 mg·L~(-1)、基本培养基类型为1/4MS、蔗糖质量浓度为15 g·L~(-1),樟叶越桔组培苗最佳生根培养基为1/4MS+IBA 2.0 mg·L~(-1)+活性炭0.1 g·L~(-1)+蔗糖15 g·L~(-1),生根率达100%,平均生根数为每株7.67条;根系呈辐射状、基部无愈伤组织,组培苗生长健壮、叶色浓绿;樟叶越桔组培苗移栽时以全腐殖土基质为佳,成活率为83.7%,植株叶片舒展,生长状况良好。该研究建立的优化体系有效地提高了樟叶越桔组培生根苗的生根率和生根质量,解决了后期移栽成活困难的问题,为优良的樟叶越桔植株规模化生产提供了科学依据和技术支持。     

模拟大气硫酸铵污染对香樟幼苗生长及光合特性的影响

通过室内盆栽实验研究了大气颗粒污染物硫酸铵对香樟幼苗生长及光合特性的影响。结果表明,香樟幼苗叶片涂抹硫酸铵处理对植物生长无显著影响;低浓度硫酸铵(2 g·L^-1)提高了叶片叶绿素含量,而高浓度(4 g·L^-1)却降低了叶片叶绿素含量;与对照相比,低浓度处理的香樟叶片净光合速率、气孔导度、胞间二氧化碳浓度与蒸腾速率无显著差异;高浓度处理的香樟叶片净光合速率与蒸腾速率高于对照,而气孔导度与胞间二氧化碳浓度与对照无显著差异。机理分析表明,硫酸铵颗粒物主要通过影响叶片气孔导度来影响植物光合特性。     

油菜愈伤组织中烯丙基硫苷的积累和某些外源碳氮化合物对其含量的影响

油菜愈伤组织在培养中生长34d的增殖量最大,增殖倍数为7．08倍,烯丙基硫苷的含量在第6天最高;外源L-缬氨酸明显促进烯丙基硫苷的积累,其浓度为2和4mmol·L^-1时烯丙基硫苷含量分别达到98．5和97．40mg·g^-1（DW）;蔗糖最有利于油菜愈伤组织的生长和其中烯丙基硫苷的积累,浓度以30g·L^-1。的蔗糖为最佳。     

茶条槭悬浮培养体系的建立与没食子酸合成的优化条件

初步建立茶条槭（Acer ginnala）细胞悬浮培养体系：以茶条槭子叶为外植体,接种于WPM培养基中,对茶条槭愈伤组织进行诱导和继代培养。悬浮培养中,每代增长指数达到11．6,没食子酸含量达到1．518％。通过对比NT、IS、WPM、B5和MS培养基所含成分对茶条槭愈伤组织悬浮培养的影响,综合考虑悬浮细胞的生长速率和有效成分的含量,确定WPM为基本培养基。WPM培养基大量元素的浓度对细胞的生长和没食子酸的积累有显著影响,其浓度越高,促进作用越明显。3倍浓度的大量元素最有利于没食子酸的积累。蔗糖浓度为10g·L^-1最适于没食子酸的积累,浓度为30g·L^-1最适于茶条槭细胞生长和没食子酸合成。     

无籽罗汉果的组织培养和快速繁殖

以无籽罗汉果优良株系的幼嫩茎段为试验材料,经消毒处理后,剪成带一个腋芽的茎段,在MS＋6-BA0．5mg·L^-1＋NAA0．05mg.L^-1培养基上进行培养,获得无菌芽苗,再以无菌芽苗的单芽茎段为外植体,建立无籽罗汉果的组培快繁体系。结果表明,最佳继代增殖培养基为MS＋6-BA0．5mg·L^-1＋IBA0．2mg·^-1＋GA30．03mg·L^-1,30d的增殖系数为16．4;芽苗伸长的最适培养基为MS＋6-BA0．05mg.L^-1＋IBA0．1mg·L^-1＋GA30．1mg·L^-1;芽苗生根的最适培养基为1／2MS＋IBA0．5mg·L^-1：炼苗后,移入蛭石：珍珠岩：熟土=1：1：2（v／v／v）的基质中,成活率达98．1％。该体系的建立为无籽罗汉果规模化生产提供了技术平台。     

‘灿烂’蓝莓组培与快繁技术

以蓝莓良种‘灿烂’半木质化茎段为材料,通过对增殖培养基以及生根培养基的筛选,建立蓝莓高效组培快繁技术体系。结果表明,最佳的增殖培养基为1/2MS＋2.0 mg·L^-1 ZT＋0.01 mg·L^-1 NAA＋30 g·L^-1白糖＋6.5 g·L^-1琼脂,芽诱导率为87%,增殖系数达3.6;ZT 1.0～2.0 mg·L^-1对不定芽诱导效果明显,随着ZT浓度的提高,愈伤组织产生越明显,影响增殖苗的质量。最佳生根培养基为1/2MS＋1.5 mg·L^-1 IBA＋0.01 mg·L^-1 NAA ＋25 g·L^-1白糖＋7.0 g·L^-1琼脂,pH值5.8,生根率70%,IBA浓度从0.5 mg·L^-1提高到1.5 mg·L^-1,生根越多,根系越壮,但也产生更多愈伤组织影响生根质量。炼苗10 d后移栽至草炭土,成活率78%。     

Photosynthesis and photoprotection in Nicotiana tabacum L. in vitro-grown plantlets

Nicotiana tabacum L. plantlets were cultured in vitro photoautotrophically (0% sucrose) and photomixotrophically (3% or 5% sucrose) at two irradiances (80 or 380 mumol m-2 s-1) with the aim of investigating the effect of these culture conditions on photosynthetic parameters and on protective systems against excess excitation energy. In plantlets grown photoautotrophically under higher irradiance photoinhibition was demonstrated. These plantlets had a decreased chlorophyll (Chl) a + b content and Chl a/b ratio, an increased content of xanthrophyll cycle pigments and a higher deepoxidation state, a decreased maximum photochemical efficiency of photosystem II (PS II) and actual photochemical efficiency of PS II, and an increased non-photochemical quenching. In the photoautotrophically grown plantlets and those photomixotrophically grown with 3% sucrose, the increase of growth irradiance from 80 to 380 mumol m-2 s-1 stimulated the activities of ascorbate-glutathione cycle enzymes with the exception of ascorbate peroxidase. Ascorbate peroxidase activity was not affected by the increase in growth irradiance but a significant decrease with increasing sucrose concentration was evident. The higher concentration of sucrose in the medium (5%) in combination with the higher irradiance inhibited photosynthesis (decrease in Chl a + b content and net photosynthetic rate) but no significant changes in activities of ascorbate-glutathione cycle enzymes were found. These results suggest that exogenous sucrose added to the medium improved high irradiance and oxidative stress resistance of the plantlets but the effect of sucrose is concentration dependent.     

外源水杨酸对NaCl胁迫下白榆幼苗光合作用及离子分配的影响

以1年生白榆幼苗为研究对象,设置0、0.5、1.0和2.0 mmol·L^-1水杨酸(SA)与0、50、100和150 mmol·L^-1 NaCl处理组合,考察盐胁迫下白榆幼苗生物量、光合色素含量、光合作用参数及根叶离子含量、分配、运输的情况,探讨外源SA对NaCl胁迫下白榆幼苗耐盐生理特征的影响。结果表明:(1)NaCl胁迫显著抑制了白榆幼苗的生长、光合色素含量及光合能力,并破坏了白榆体内离子平衡。(2)喷施外源SA使盐胁迫下白榆幼苗的干重和根冠比均不同程度升高,0.5和2.0 mmol·L^-1 SA不同程度提高了50和100 mmol·L^-1 NaCl处理组幼苗叶片光合色素含量。(3)0.5 mmol·L^-1 SA显著提升了50 mmol·L^-1 NaCl处理组白榆幼苗的净光合速率(Pn)、气孔导度(Gs)和蒸腾速率(Tr),1.0和2.0 mmol·L^-1 SA对150 mmol·L^-1 NaCl处理组幼苗净光合速率改善效果较好,外源SA对100 mmol·L^-1 NaCl处理组幼苗的光合作用参数无显著影响。(4)NaCl胁迫下,外源SA处理的白榆幼苗叶和根Na^+含量及Na^+/K^+、Na^+/Ca^2+和Na^+/Mg^2+显著降低,离子选择运输系数SK,Na、SCa,Na和SMg,Na升高,从而促进了幼苗K^+、Ca^2+和Mg^2+由根向叶片的转运;隶属函数分析发现对白榆幼苗叶和根中离子含量改善效果最好的SA浓度分别为1.0和2.0 mmol·L^-1。因此,适宜浓度的外源水杨酸能够有效改善NaCl胁迫下白榆幼苗的光合能力,有效调节白榆幼苗体内离子状态,从而增强白榆对NaCl胁迫的抗性。     

连作土壤中施加球毛壳ND35菌肥对杨树生理特性的影响

在山东农业大学林学实验站杨树连作土壤中施加不同剂量(0.17、0.33、0.67、1.00、1.33和1.67 g·kg^-1)的球毛壳ND35菌肥,测定不同处理下一年生杨树叶片的光响应过程、叶绿素荧光以及叶黄素循环等光合生理生化指标,研究球毛壳ND35菌肥对杨树根系和光合生理性能的影响.结果表明: 随连作土壤中菌肥含量的增加,杨树叶片的叶绿素含量(Chl)呈增加趋势,电子传递速率(ETR)、净光合速率(P_n)、量子效率(Φ)、硝酸还原酶(NR)活性以及根系活力等生理指标均呈先增加后降低的趋势,依赖于叶黄素循环的光合热耗散呈降低趋势,而非光化学淬灭(NPQ)呈先降低后增加的趋势.在土壤施加菌肥剂量0.67和1.00 g·kg^-1处理时,光合机构的过剩光能减少,向光化学反应方向分配的光能增多,光能利用效率提高.在连作土壤中施加适量(0.67～1.00 g·kg^-1)的球毛壳ND35菌肥,能在一定程度上提高杨树的根系生理活性,提高杨树叶片对光能的利用效率,有利于改善杨树叶片的光合机构运转状态,提高叶片的光合作用效率.     

Reduction of ribulose-1,5-bisphosphate carboxylase/oxygenase efficiency by the presence of sucrose during the tissue culture of strawberry plantlets

Summary In order to identify the physiological and biochemical events leading to the negative effects of the presence of sucrose in culture medium on the photosynthetic capacity of plantlets cultivated in vitro, time course in photosynthesis, metabolite pool sizes, and ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.39) activity were investigated in strawberry (Fragaria x ananassa Duch. cv. Kent) plantlets following their transfer to medium with or without sucrose. When the plantlets grown in medium without sucrose were transferred to a similar medium with 30 g liter⁻¹ sucrose, their net photosynthesis decreased and their level of phosphorylated compounds increased with time. In addition, initial catalytic turnover, total catalytic turnover, and the activation state of ribulose-1,5-bisphosphate carboxylase decreased in these plantlets. Conversely, when the plantlets grown in medium with 30 g liter⁻¹ sucrose were transferred to a similar medium without sucrose, their net photosynthesis slowly increased with time and their level of phosphorylated compounds slowly decreased. A slow increase with time of initial catalytic turnover, total catalytic turnover, and the activation state of ribulose-1,5-bisphosphate carboxylase was also observed in these plantlets. The results of the present paper suggest that the reduced photosynthetic capacity of strawberry plantlets cultivated in vitro in the presence of sucrose is the consequence of a reduction in the efficiency of ribulose-1,5-bisphosphate carboxylase/oxygenase due to its deactivation and the possible presence of putative inhibitors of carboxylation sites.     

Similarity of growth patterns between plantlets and seedlings of Brassica campestris L. under different in vitro environmental conditions

Explants and seeds of Brassica campestris L. were cultured on Murashige & Skoog (1962) medium with and without sucrose in a vessel with different numbers of air changes per hour under different PPF (photosynthetic photon flux) conditions. The growth and development of plantlets in the vessel were similar to those of seedlings when cultured under the same in vitro environmental conditions. The growth and development of seedlings when cultured under the same in vitro environmental conditions. The growth and development of plantlets/seedlings were greater for treatments with a higher number of air changes per hour and a higher PPF regardless of the sucrose concentration in the culture medium.The CO₂ concentration in the vessel with a lower number of air changes per hour decreased to approximately 100 ppm during the photoperiod on day 21 due to the photosynthetic activities of the plantlets/seedlings. The low CO₂ concentration, in turn, reduced the net photosynthetic rate of plantlets/seedlings in the vessel, and thus affected their growth and development.Abbreviations C_in CO₂ concentration in the culture vessel - C_out CO₂ concentration in the culture room - MS mineral composition of Murashige & Skoog (1962) medium - PPF photosynthetic photon flux     

大薯类原球茎的离体诱导及再生体系的建立

采用正交试验设计方法,以大薯带节茎段为外植体,离体诱导类原球茎并建立大薯类原球茎的再生体系,以解决愈伤组织分化成苗和试管苗移栽成活率低的难题。结果表明：以带节茎段为外植体诱导类原球茎的最适培养基为MS（含3×Ca2＋）＋1.0 mg·L-1 6-BA＋0.2 mg·L-1 NAA＋0.1%PVP＋3%蔗糖,诱导率高达93.33%;类原球茎增殖的最适培养基为MS＋4mg·L-1 6-BA＋80 mg·L-1 Ad＋0.1%PVP＋3%蔗糖;类原球茎生根的最适培养基：1/2MS＋0.10 mg·L-1 NAA＋0.1%PVP＋3%蔗糖。将诱导得到的生根类原球茎植株进行炼苗,移栽基质珍珠岩：蛭石=2：1,移栽成活率可达到95%。     

NaCl胁迫对萝卜幼苗叶片生理特征及根尖细胞核形态的影响

对0.00(对照)～0.50 mol·L-1NaCl处理48 h后萝卜(Raphanus sativus L.)幼苗叶片的叶绿素含量、净光合速率、丙二醛(MDA)含量和相对电导率进行了测定,并观察了根尖细胞核形态变化和胁迫后幼苗生长的恢复状况.测定结果表明:经0.05 mol·L-1NaCl处理后,叶片的叶绿素含量和净光合速率均高于对照(0.00 mol·L-1 NaGl);而用0.10～0.50mol·L-1NaCl处理后,叶绿素含量和净光合速率均随NaCl浓度的提高逐渐降低且与对照差异显著,其中,经0.35 ～ 0.50 mol·L-1 NaCl胁迫处理后叶片净光合速率降至0.00μmol·m-2·s-1.随NaCl浓度的提高,MDA含量和相对电导率呈先快速上升而后逐渐下降的趋势,且均与对照差异显著;其中,经0.30 mol·L-1 NaCl处理后MDA含量最高,经0.40 mol·L-1NaCl处理后相对电导率最高.观察结果显示:经0.05～0.25 mol·L-1NaCl处理后根尖细胞核由规则的圆形向各种不规则形状转变,染色质出现不同程度的浓缩,表现出不同程度的程序性细胞死亡特征;经0.30 ～ 0.50 mol·L-1NaCl处理后则出现中空细胞与部分正常细胞并存的现象.经0.05和0.10 mol·L-1 NaCl处理后,萝卜幼苗可恢复正常生长,且株高与对照无明显差异;而经0.25 ～ 0.50 mol·L-1 NaCl处理后,幼苗均较难恢复正常生长,且随NaCl浓度的提高,幼苗出现枯萎、糜烂现象.研究结果显示:低浓度NaCl处理不仅对萝卜幼苗的生长、生理代谢及细胞核形态结构没有明显伤害反而有一定的促进作用,而高浓度NaCl胁迫可导致萝卜幼苗不可逆伤害.     

Culture on sugar medium enhances photosynthetic capacity and high light resistance of plantlets grown in vitro

The significance of photosynthetic photon flux (PPF) and sugar feeding for the production of plants in vitro is only poorly understood. Nicotiana tabacum L. plantlets were grown photoautotrophically and photomixotrophically (3% sucrose) at two different PPFs (60 µmol m⁻² s⁻¹ and 200 µmol m⁻² s⁻¹) to investigate the effect of these culture parameters on photosynthetic performance and growth. Photomixotrophically‐grown plantlets showed an increase in carbohydrate content, mainly in glucose and fructose. Plant growth, dry matter accumulation and total leaf area were higher under photomixotrophic than photoautotrophic conditions. Not only biomass formation but also photosynthesis was positively affected by exogenous sucrose; the chlorophyll (Chl) content and the light‐saturated rate of photosynthetic oxygen evolution were higher in photomixotrophic plantlets. Photoinhibition occurred in plantlets that were grown photoautotrophically at the higher PPF. It became apparent as a loss in Chl content and photochemical efficiency. Photoinhibited plantlets showed a decrease in the D2/LHCII and CP47/LHCII ratios, suggesting a preferential loss of proteins from the photosystem II (PSII) core. The increased content of xanthophyll cycle pigments in photoinhibited plantlets indicated that also protective mechanisms were activated. Photomixotrophic growth of the plantlets prevented the occurrence of photoinhibitory symptoms. Therefore, we conclude that culture on sugar medium increases not only the photosynthetic potential but also the high light resistance of plantlets grown in vitro.     

翅梗石斛组织培养与快繁技术(简报)

以云南野生翅梗石斛Dendrobium trigonopus种子无菌萌发的试管苗为材料,初步探讨翅梗石斛的组织培养和快速繁殖技术。结果表明：B5 + NAA 0.5 mg·L^-1 + 10%马铃薯汁和B5 + NAA 0.5 mg·L^-1 + 6-BA 0.5 mg·L^-1 + 10%马铃薯汁培养基分别对翅梗石斛种子的萌发与原球茎的增殖效果明显;B5 + NAA 0.5 mg·L^-1 + 10%马铃薯汁+云南松树皮粉末2 g·L^-1培养基对翅梗石斛的生根及壮苗效果较好。成苗后进行简单的药剂灭菌,便可直接移栽至锯末与刨花或锯末与树皮比例为1∶1的基质中,25 d后,成活率达95%以上。