不同生境条件对小蓬竹生长的影响

小蓬竹具有较高的经济、社会和生态价值,且为濒危竹种,但繁殖成活率很低,引种栽培长势也较原生境差。为探索上述原由,在野外样地调查的基础上,利用SPSS软件进行相关性分析,研究小蓬竹在不同生境条件下的生长状况,为小蓬竹的经营、利用和保护提供科学依据,结果表明：①小蓬竹生长受坡向影响很大;②坡位对小蓬竹生长的影响并不大;③小蓬竹在不同坡度均能够生长,坡度30＆#176;以下较利于小蓬竹生长;④海拔对小蓬竹的生长有很大的影响。     

Mevalonate kinase activity during different stages of plant regeneration from nodular callus cultures in white pine (Pinus strobus)

Mevalonate kinase (MK) catalyzes a step in the isoprenoid biosynthetic pathway, which leads to a huge number of compounds that play important roles in plant growth and development. Here, we report on changes in MK activity in white pine (Pinus strobus L.) during plant regeneration by adventitious shoot organogenesis from cotyledons of mature embryos, including nodular callus induction, shoot formation and rooting. Nodular calli were induced from Pinus strobus (PS) embryos by culture in nodular callus induction medium in a 0-, 8- or 16-h photoperiod. Mevalonate kinase activity peaked in nodular calli after three weeks of culture on nodular callus induction medium in a 16-h photoperiod, whereas frequency of nodular callus formation peaked after 4 weeks of culture on nodular callus induction medium in darkness. During adventitious shoot formation, MK activity peaked in shoots derived from dark-grown nodular calli after 3 weeks on bud formation medium, and frequency of shoot formation was highest in dark-grown nodular calli cultured on bud formation medium for 4 weeks. During rooting, MK activity peaked 2 weeks after transfer of adventitious shoots to rooting medium and rooting frequency was highest in adventitious shoots after 3 weeks on rooting medium. Although during nodular callus induction in darkness MK activity was inversely related to frequency of nodular callus formation, MK activity was highly correlated with frequency of shoot formation and with rooting frequency. The observed increase in MK activity preceding rooting suggests that MK could serve as a marker for rooting of white pine shoots in vitro.     

印度橡皮树愈伤组织培养成苗技术

以印度橡皮树顶芽和茎段为材料,进行组织培养试验,结果表明:脱分化培养基最佳配方MS+6-BA 2 mg/L+NAA 0.05 mg/L+2,4-D 0.25 mg/L;继代培养基最佳配方:MS+6-BA 5 mg/L+NAA 0.02mg/L;生根培养基最佳配方:MS+IBA 0.5mg/L。印度橡皮树的炼苗生根成活率一般都在95%以上。     

油松成熟胚的不定芽诱导及植株再生

以油松成熟合子胚为外植体,通过器官直接发生途径获得再生植株。诱导油松不定芽的最佳基本培养基为DCR,最佳激素组合为TDZ0.1mg/L+NAA0.2mg/L,不定芽生长的最佳激素组合为TDZ0.05mg/L+NAA1.0mg/L;生根最佳培养基为1/2DCR+IBA2mg/L+6-BA0.05mg/L。生根苗移栽到草炭土∶沙=2∶1(体积比)的基质中,成活率为80%。     

“突尼斯软籽”石榴叶片愈伤组织再生体系的建立

为了石榴遗传转化研究的开展,以突尼斯软籽石榴为研究材料,叶片作为外植体,研究了不同灭菌时间和不同生长调节物质对离体叶片愈伤组织再生的影响。结果表明:最适宜的暗培养时间为14 d,最佳愈伤组织分化培养基为MS+BA 1.0 mg.L-1+NAA(0.3～0.5)mg.L-1,最佳不定芽分化培养基为MS+NAA 0.3 mg L-1+TDZ 1.5 mg L-1+KT(1.5～2.0)mg L-1,最佳新梢增高的培养基为MS+NAA 1.0 mg L-1+BA 0.2 mg L-1+KT 1.0 mg L-1,适宜生根的培养基为MS+NAA0.5 mg L-1,生根率达100%。     

Changes in cellular structures and enzymatic activities during browning of Scots pine callus derived from mature buds

Visible browning is a typical feature of callus cultures derived from shoot tips of mature Scots pine (Pinus sylvestris L.). Because the ability of callus to regenerate is low, we determined the effect of browning on growth and changes in cellular structure during culture. Striking alterations in cellular structure were detected by LM (light microscopy), EM (electron microscopy) and SEM (scanning electron microscopy). Accumulation of phenolic substances was shown by histochemical staining. Staining for beta-glucosidase activity of soluble proteins that had been subjected to polyacrylamide gel electrophoresis indicated lignification of cells. The measured growth rate of callus was low compared with a hypothetical growth curve. Peroxidase activity increased rapidly soon after the start of the culture period, but especially between the second and third weeks of culture. At this time, the degradation of cell membranes and browning began coincident with the loss of chlorophyll. We conclude that browning is associated with cell disorganization and eventual cell death, making tissue culture of mature pine especially difficult.     

Efficient embryogenic callus induction and plant regeneration from embryonic axis explants inQuercus acutissima

Embryogenic callus ofQuercus acutissima was successfully induced from embryogenic cultures, and plants were regenerated from the callus. The development of the techniques involved will allow mass propagation and gene transformation in this species. Embryogenic cultures were formed from embryonic axis explants (i.e., embryos without cotyledons) excised from immature embryos, after culture on Murashige and Skoog (MS) medium containing indolebutyric acid and benzyladenine. Attempts to induce embryogenic cultures from cotyledon explants were unsuccessful. Embryogenic calli were induced at high frequency from embryogenic cultures on MS medium containing 2,4-dichlorophenoxyacetic acid. However, benzyladenine inhibited embryogenic callus formation. Somatic embryo development from embryogenic calli occurred on MS medium in all of the seven cell lines tested. Germination of somatic embryos was induced on half strength MS medium without plant growth regulators. Finally, acclimated plants growing in soil were obtained.     

毛白杨叶片愈伤组织的诱导、芽的分化及不定根的再生

以毛白杨叶片为外植体进行组织培养研究。结果表明:培养基MS 6-BA 1.0 mg/L NAA 1.0 mg/L有利于愈伤组织的诱导;培养基MS 6-BA 0.5 mg/L NAA 0.5 mg/L诱导愈伤组织并且有大量不定芽发生;培养基MS ZT 0.1 mg/L NAA 0.3 mg/L可使叶片生根率达100%。     

花曲柳茎芽增殖和植株再生

为建立高效的花曲柳组培快繁体系,以花曲柳休眠芽萌发获得的带芽茎段和成熟种子无菌萌发获得的带芽茎段为外植体,通过基本培养基和不同植物生长调节剂组合的筛选,诱导出丛生芽,获得了再生植株.结果表明:花曲柳休眠芽采集的最适时期是4月中旬,适合的基本培养基为改良的B5培养基,在改良的B5培养基上添加BA 4.0 mg·L-1和I...     

Plant regeneration of transgenic China Rose (Rosa chinensis Jacq.) from organogenic callus

Different types of explants of China Rose (Rosa chinensis Jacq.) were placed on a Schenk and Hildebrandt (SH) medium containing L-proline and 2,4-dichlorophenoxyacetic acid (2,4-D). Organogenesis was observed on callus induced from both whole leaf and petiole and the high frequency of organogenesis was observed on the whole leaf. Shoot regeneration was obtained via organogenesis. The effects of pH and concentrations of antibiotics on maintenance of organogenesis capacity were investigated in subsequent subcultures. The pH value was found to play a critical role in retaining organogenesis capacity. The binary vector pBI121, carrying the gus gene coding forβ-glucuronidase (GUS) and the nptⅡgene mediated by Agrobacterium tumefaciens, was used for transformation of organogenic callus using 50 mg·L-1 geneticin for selection. Six regenerated lines showed GUS activity, of which five were verified for the presence of nptⅡgene by PCR.     

邻体干扰改进模型在竹类研究中的利用——以小蓬竹为例

依据改进的邻体干扰模型,利用喀斯特适生竹种小蓬竹样地实际调查的基株与邻体的数据,研究基株竹高、冠幅以及枝下高与干扰指数的关系,结果表明:基株的冠幅、树高与枝下高与干扰指数之间呈显著的负相关关系。不同样地因为密度不同,相同因子相同数值决定的干扰指数不一样。该研究结果为合理营造和经营竹类林分提供理论依据。     

Somatic Embryogenesis and Plantlet Regeneration in Callus Cultures Derived from Mature Zygotic Embryos of Slash Pine

1TheworkwassupportedbyChineseNationalHighTechnologyDevelopmentProgram"863project"IntroductionSomaticembryogenesishasagreatpotentiaIforrapidProPaationofsuPCriorconifcrspccics.ThefirstreportonsomaticembryogenesisandplantlctrcgencrationinconifersPeCieswasach…     

In vitro plant regeneration from cotyledon-derived callus cultures of leguminous tree Gleditsia caspica Desf.

An in vitro regeneration system was developed using organogenic callus derived from in vitro grown cotyledonary explants of Gleditsia caspica Desf., an important leguminous tree. Murashige and Skoog (MS) basal medium augmented with 0.2 g L^?1 myo-inositol and various concentrations of either 2,4-dichlorophenoxyacetic acid (2,4-D), naphthaleneacetic acid, or indole-3-butyric acid (IBA) alone as well as combined with cytokinins was used for callus induction. The highest frequency of organogenic yellowish-white and nodular callus (93 %) was obtained from explants grown on medium supplemented with 13.5 μM 2,4-D and 4.4 μM benzyladenine (BA). The yellowish-white and nodular callus when transferred to MS medium supplemented with BA (2.2–17.7 μM) or kinetin (KT; 2.3–18.8 μM) solely or in combination with 2.3 μM 2,4-D produced several microshoots after 5 weeks culture. The calli cultured on MS medium with 4.4 μM BA singly showed superior growth response and produced both maximum shoot regeneration (94 %) and the highest mean number (4.3) of microshoots per callus. Transfer of regenerated microshoots onto modified MS basal medium fortified with 5.8 μM gibberellic acid and 4.4 μM BA resulted in the maximum number of internodes per shoot and the highest shoot elongation after a period of 6 weeks. Optimum rooting of 90 %, an average 6.1 roots per shoot, and a mean root length of 3.6 cm was observed when half-strength MS medium was supplemented with 9.8 μM IBA and 0.92 μM KT. The regenerated healthy plants with well-developed shoots and roots showed a survival rate of 77 % after acclimatization and transplanting to garden soil for a 10-week hardening period under ex vitro conditions.     

紫穗槐茎段愈伤组织诱导及再生体系的建立

以紫穗槐茎段为研究对象,确定了影响其愈伤组织诱导的关键因子、不定芽分化及生根培养的最佳条件,建立了紫穗槐稳定高频再生体系。试验结果表明：紫穗槐茎段愈伤组织的最适诱导培养基为MS＋6-BA4.0mg/L＋NAA2.0mg/L＋2,4-D0.5mg/L,诱导率为100%,经愈伤组织分化不定芽的最佳培养基为MS＋6-BA0.5mg/L＋NAA0.1mg/L＋KT2mg/L,分化率为96.2%,增殖倍数为7.3;不定芽最佳生根培养基为1/2MS＋酵母提取物0.5mg/L;再生植株移植在选用田园土、蛭石（5∶1）混合的基质上,光强3000lx,光照时间14h/d下生长,3周后成活率达85.67%。     

草食蚕胚性愈伤组织的诱导及再生体系的建立

为给草食蚕的快繁研究提供参考,以草食蚕无菌苗为材料,比较研究不同激素配比条件下不同外植体愈伤组织的诱导及植株再生。结果表明,草食蚕无菌苗叶片、叶柄和茎段均可作为离体培养的外植体,但叶柄效果最好。将外植体接种于添加6-BA(0.3～1.5mg/L)和NAA(0.05～0.80mg/L)的MS培养基上,经14～16d培养,可100%脱分化产生绿色致密愈伤组织,且愈伤组织分化产生大量不定芽。将单芽茎段接种于添加KT(0.1～1.0mg/L)和NAA(0.05mg/L)的MS培养基上,经4周培养,产生的芽苗健壮,增殖系数高达7.86。将高约3cm的芽接种于分别添加IAA(0.1～0.5mg/L)、IBA(0.1～0.5mg/L)或NAA(0.05～0.10mg/L)的1/2MS培养基上,可100%产生不定根,获得完整的再生植株。生根苗采用"二步法"移栽,移栽成活率为92.5%。     

Indirect somatic embryogenesis and regeneration of Fraxinus mandshurica plants via callus tissue

Somatic embryogenesis of Fraxinus mandshurica has the problems of low somatic embryo(SE) yield,unsynchronized SE development,and a high percentage of deformed SEs.We aimed to improve F.mandshurica SE production by synchronizing SE development,improving SE quality,and inducing root formation to obtain complete regenerated plants.Cotyledons of immature zygotic embryos of F.mandshurica were induced to form callus and then SEs.The SE induction percentage from explants differed among 32 mother trees,and the one with the highest SE induction percentage(29.8%) was used for further experiments.The highest callus induction percentage was94.2% on 1/2-strength Murashige and Skoog medium(MS1/2)supplemented with 0.15 mg·L~(-1) naphthalene acetic acid.The highest callus proliferation coefficient(240.5) was obtained on McCown's Woody Plant Medium containing 0.1 mg·L~(-1)6-benzyl adenine and 0.15 mg·L~(-1) 2,4-dichlorophenoxyacetic acid.The highest number of SEs(1020.5 g~(-1) fresh weight) was obtained on MS1/2 medium supplemented with1 mg·L~(-1) 6-benzyladenine.The highest number of cotyledon embryos(397/g fresh weight) was obtained by incubating materials on medium containing 1 mg·L~(-1) abscisic acid and then applying a drying treatment.The cotyledon embryos were milky white,uniformly sized(average length 4.7 mm),and 80% of them were normal.The SE rooting percentage on 1/2 MS medium containing 0.01 mg·L~(-1) NAA was 37.5%.Overall,the germination percentage of SEs was 26.4%,and complete regenerated plants were obtained after transplanting and acclimation.These results provide more possibilities for the preservation and breeding of F.mandshurica.     

Selection, regeneration and protein profile characteristics of NaCl-tolerant callus ofRobinia pseudoacacia L.

Changes in callus growth, differentiation potency and protein profile induced by salt stress were investigated in the calli induced from a hypocotyl ofRobinia pseudoacacia L. The NaCl treatment evidently obstructed the callus growth and the influences seemed to continue to be present in the selected calli even after they were restored in NaCl-free medium that could partially recover the callus growth, and some polypeptides were found to restore. The calli selected and restored intermittently every two weeks for one year completely lost their differentiation potency. The NaCl-tolerance test of the regenerated plantlets revealed that part of the plantlets that directly regenerated from the selected NaCl-tolerant calli were able to survive in 0.2 M NaCl, while most of the plantlets regenerated from the NaCl-nonselected calli or from restored calli were obviously damaged by this treatment. The SDS-PAGE assay indicated that a 54 kDa polypeptide increased distinctly to a very high level while a 41 kDa polypeptide decreased to a lower level in almost all the NaCl-tolerant plantlets. This observation led to the suggestion that the changes may play an adaptive role in allowing plantlets to survive under saline conditions.     

东部白松试管芽诱导和增殖技术研究

以东部白松种子和幼嫩顶芽为外植体,研究影响试管芽获取的关键因子、增殖的最佳培养基种类及添加激素种类和浓度组合,有效建立了东部白松无菌试管芽增殖体系。结果表明：以0～4℃沙藏6周的东部白松种子为外植体,用0．I％HgCl2消毒10min,获得了无菌试管芽,试管芽的最佳增殖培养基为B5＋ZT 0．6mg／L＋IBA 0．4mg／L,增殖系数达4．09。     

Histological studies of shoot regeneration system in hypocotyl-derived callus of Phellodendron amurense Rupr.

An efficient in vitro plant regeneration was achieved from hypocotyl-derived callus of a medicinal tree Phellodendron amurense. The expected morphogenic response was obtained on the media containing 0.89 µM 6-benzyl aminopurine (BAP) plus 4.52 µM 2.4-dichlorophenoxyacetic acid (2,4-D), 4.44 µM BAP plus 5.37 µM α-naphthaleneacetic acid (NAA), or 2.22 µM BAP plus 4.92 µM indole-3-butyric acid (IBA). A detailed histological study was undertaken to gain a better understanding of cellular changes that take place during the plant regeneration process from callus tissue. This study led to the identification of the cellular origin of shoot regeneration. Histological studies revealed that the new vegetative buds originated from callus that completely altered the morphology of the callus tissues by the 60th day of culture. It also revealed that BAP with NAA or IBA induces plant regeneration through organogenesis whereas BAP with 2,4-D induces embryo-like-structure (ELS) through hypocotyl-derived callus. The presence of ELS lacking root poles was also observed.     

Selection of culture conditions for callus induction and proliferation by somatic embryogenesis of Pinus koraiensis

The induction and proliferation of embryogenic callus are key steps for large-scale propagation of somatic embryogenesis pathway and long-term preservation of coniferous germplasm.Callus can be induced from immature embryos of Korean pine(Pinus koraiensis Sieb.et Zucc.;Pinaceae) as explants,but there are problems,such as low proliferation efficiency,loss of embryogenicity,poor vigor;thus,best conditions for proliferation and culture of immature embryos of Korean pine are not yet clear.To solve the problems with somatic embryogenesis of Korean pine and determine the best culture conditions for callus induction and proliferation,we varied hormone concentration,subculture cycle of proliferation and other plant growth regulators combinations in media to induce callus formation by megagametophytes of three Korean pine families at different developmental stages,then analyzed the effects on embryogenic callus retention and cell proliferation using a quadratic regression orthogonal rotation design.The results showed that the family origin and collection date of explants significantly affected callus induction(induction rate reached1.67%).Embryogenic maintenance and callus proliferation were best on DCR medium supplemented with 0.25 mg L~(-1)6-benzyl adenine,1 mg L~(-1) naphthaleneacetic acid,30 g L~(-1)sucrose,500 mg L~(-1),L-glutamine,500 mg L~(-1) casein hydrolysis and 6.5 g L~(-1) agar.In addition,the combination of 2,4-dichlorophenoxyacetic acid+6-benzyl adenine also had a better proliferative effect on callus.The effects of different combinations of growth regulators on callus proliferation efficiency were significantly different.Transfer to new medium every 13-15 days not only maintained robust callus vigor,but also yielded a larger proliferation coefficient.The techniques and conditions for embryogenic callus induction and proliferation of Korean determined here will serve as a foundation for establishing a large-scale system for somatic embryogenesis and propagation of Korean pine.