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。     

铁核桃离体培养与快速繁殖

以优良单株‘纳雍-1’的单芽茎段为外植体,建立了铁核桃（Juglanssigillata）离体培养与快速繁殖的体系。结果表明,附加6-BA1．0mg·L-1 ＋活·IgK（AC）3．0g·L-1的DKw培养基适宜铁核桃腋芽诱导;适宜铁核桃芽增殖的培养基为DKW＋6-BA1．0mg·L-1 ＋IBA0．02mg·L-1,40d后增殖系数可达7．33;试管苗的茎尖和茎段均可用于增殖培养;一步生根法（低浓度的生长素IBA持续诱导）不利于铁核桃试管苗嫩茎生根;采用二步生根法,生根率最高可达71．73％,其中,不同IBA浓度、暗培养时间、蔗糖浓度和AC含量对试管苗嫩茎生根影响显著,铁核桃试管苗在附]sulBA5,0mg·L-1的1／4DKW培养基中暗培养12d,再转移到不含IBA的1／4DKW培养基（附加AC 3g-L-1和蔗糖20g·L-1）中生根效果最好;生根试管苗采用珍珠岩和营养土两步炼苗,60d后成活率达到87．50％。     

香果树茎段和叶片的组织培养

以香果树（Emmenopterys henryi Oliv．）实生苗带芽茎段和叶片为外植体进行组织培养。结果表明,香果树带芽茎段愈伤组织的最适诱导培养基为含有1．0mg·L^-1～6-BA和0．01mg·L^-1 NAA的MS培养基,愈伤组织诱导率达100％;愈伤组织分化的最适培养基为添加2．0mg·L^-1KT和0．01mg·L^-1 NAA的MS培养基;在含有1．0mg．L^-1～6-BA和0．1mg·L^-1 NAA的MS培养基上,叶片诱导不定芽的效果较好,诱导率可达80％;在含有2．0mg．L^-1 KT和0．1mg·L^-1 NAA的MS培养基上,不定芽的增殖系数可达3—4;以含有1．5mg·L^-1 IBA的1／2MS培养基为生根培养基,香果树试管苗生根率达72．73％。移栽至大田后,香果树试管苗的成活率达到30％。     

‘巴斗’杏再生体系的建立与耐盐突变体的筛选

以‘巴斗’杏试管苗茎段为外植体,研究其再生体系的建立以及在含不同浓度NaCl培养基上诱导耐盐愈伤组织,筛选耐盐突变体。结果显示：茎段在MS＋6-BA1．5mg·L^-1＋IBA0．5mg·L^-1培养基上诱导愈伤组织效果最好,芽的分化率可达88％;将出芽愈伤组织块接种到附加IBA0．5mg·L^-1＋KT2．0mg·L^-1的MS分化培养基上效果最佳,芽的分化系数最高为12．7;较理想的生根培养基为MS＋NAA0．1mg·L^-1。＋IBA0．2mg·L^-1,生根率在46．3％以上;在含0．8％NaCl的愈伤组织诱导培养基中,连续继代筛选2代,转入不含NaCl的分化培养基中,分化出了完整植株。经继代培养筛选,测定发现获得的耐盐植株比正常培养植株的游离脯氨酸含量高。     

石龙尾(Limnophila sessiliflora BI.)的组织培养与快速繁殖技术研究

以石龙尾（Limnophila sessiliflora BI．）沉水枝带节茎段为外植体进行离体培养,研究外植体灭菌方法以及培养基中不同生长调节剂的浓度对其增殖、生根的影响。结果表明：以0．1％的HgCl2为灭菌剂,采用4min＋4min、间歇4h的间歇灭菌法,可以获得成活的无菌外植体15％;在1／2MS＋6．BA2．0mg·L^-1＋NAA 0．1～0．2mg·L^-1的增殖培养基上培养35d,试管苗的增殖系数可达30．8以上;在1／2MS＋6-BA 0．3mg·L^-1＋NAA 0．5mg·L^-1的生根培养基上培养28d后,可获得具3～5个侧枝的生根苗,平均每株生根数4．8条;炼苗后移植成活率100％。     

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。     

米老排的组织培养和快速繁殖

以米老排人工林成年优树当年生枝条茎段为外植体,建立了“以芽繁芽”的组织培养快速繁殖体系。丛芽诱导培养最快的无性系,经过3个月的培养,一个外植体可获得17．2个芽。在添加6-BA1．0mg·L-1的Ms培养基上增殖率最高,月增殖率为2．43。促进苗高生长的最有效培养基是Ms＋6-BA0．4mg·L-1＋GA30．4mg·L-1。生根培养基为1／2MS＋IBA0．4nag·L-1＋O．1g．L-1活性炭,生根率达81．8％以上,每株苗生根7．8条,平均苗高为1．2cm。经生根培养20d和目光温室炼苗15d后,试管苗移植入黄泥和泥炭土（4：1,刃功混合基质中,成活率达85％以上。     

影响芋茎尖培养中脱毒和快速繁殖的几个生理因素

以3个芋品种（‘石川早生’、‘虾籽芋’、‘叶用芋）球茎茎尖为外植体,进行脱病毒和快繁的结果表明,外植体表面灭菌的最佳方法是剥鳞片→乙醇→新洁尔灭→剥幼叶→氯化汞;适宜茎尖分化的培养基为MS＋1．0-2．0mg·L^-16-BA＋0．2mg·L^-1 NAA。生物学方法和电镜观察显示：连续3代0．5-0．7mm茎尖剥离培养对芋花叶病毒（DMV）的脱毒率达100％。在培养基MS＋0．2mg·L^-1 NAA中,适量添加6-BA和TDZ,三品种芋的试管苗增殖效果好;附加KT,试管苗生长健壮且利于生根：添加20-100mg·L^-1的精胺（spm）,可促进不定芽的发生,与KT配合使用可促使继代增殖和成苗一步完成。完整植株在草炭土：蛭石=1：1的基质中,移栽成活率超过97％,且苗生长健壮。     

油茶良种‘华硕’的组织培养及高效生根

以油茶‘华硕’带芽茎段为外植体,研究植物生长调节剂、珍珠岩对其快速繁殖及试管苗生根的影响。结果表明：茎段腋芽萌发的最佳培养基为：MS＋2.0 mg·L^-1 6-BA＋0.1 mg·L^-1 IAA,萌发率达88.68%;最佳继代增殖培养基为：WPM＋3.0 mg·L^-1 6-BA＋0.01 mg·L^-1 IBA＋6.0 mg·L^-1 GA3,增殖系数可达11.27;最佳壮苗伸长培养基为：WPM＋0.05 mg·L^-1 IAA＋6.0 mg·L-1GA3;最佳生根培养基为：1/2MS＋1.0 mg·L^-1 IBA＋50 g·L^-1珍珠岩,生根率95.83%。炼苗后移栽到泥炭土、珍珠岩、黄土（1：1：1,V/V/V）混合基质中,成活率达85%以上。     

‘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%。     

Effect of agar concentration on growth and anatomy of adventitious shoots of Picea abies (L.) Karst

The development of adventitious shoots of Picea abies was affected by the agar concentration in the culture medium. Increasing the agar concentration from 0.5 to 2.0% decreased vitrification, but at the same time reduced shoot growth and rooting potential. Slightly vitrified plantlets could become acclimatized to greenhouse conditions. The mesophyll of needles developed in vitro was interspersed with large air spaces; the lower the agar concentration, the larger the air spaces. After transfer to the greenhouse, the new needles from the acclimatized plantlets had an anatomy approaching that of plants growing in field.     

Supporting material affects the growth and development of in vitro sweet potato plantlets cultured photoautotrophically

Summary A comparative study was conducted to optimize the vegetative growth of sweet potato (Ipomoea batatas L. (Lam), cv. Beniazuma) plantlets cultured in vitro in five different types of supporting materials: agar matrix (a seaweed derivative; Kanto Chemical Co. Inc., Tokyo), gellan gum (a Pseudomonas derivative; Kanto Chemical Co. Inc., Tokyo), vermiculite (a kind of hydrous silicates), a mixture of vermiculite and cellulose fiber (Florialite; Nisshinbo Industries, Inc., Tokyo) and cellulose plug (Sorbarod; Baumgartner Rapiers SA., Switzerland). Single nodal cuttings were cultured photoautotrophically (without any sugar in the medium and with enriched CO₂ and high photosynthetic photon flux) for 21 d on MS basal medium. Plantlets exhibited the greatest growth when Florialite was used as supporting material. The leaf and root fresh and dry mass were 2.4× and 2.9×, and 2.2× and 2.8× greater, respectively, than those of the plantlets grown in the agar matrix (control). Plantlets cultured in Sorbarod supporting material exhibited the second greatest fresh and dry mass of leaves and roots followed by vermiculite and gellan gum supporting material. The most interesting feature was the development of a large number of fine lateral roots from the main adventitious root in the Florialite treatment. Among the treatments, the highest net photosynthetic rate was evident in the Florialite grown plantlets. The percent porosity of the supporting materials was highest in Sorbarod followed by Florialite and vermiculite. Plantlets transplanted from the Florialite supporting material exhibited the highest acclimatization percentage followed by that of the Sorbarod treatment.     

Micropropagation of Terminalia arjuna Roxb. from cotyledonary nodes

Cotyledonary node explants excised from 21 day old seedlings of T. arjuna produced multiple shoots when cultured on full strength MS or modified MS (1/2 strength major salts and Fe-EDTA) medium supplemented with different concentrations (0.1-1.0 mg/l) of BAP. Maximum 8.9 shoots/explant could be recorded after 30 days of inoculation on modified MS medium supplemented with BAP (0.5 mg/l). A proliferating shoot culture was established by reculturing the original cotyledonary nodes (2-3 times) on shoot multiplication medium after each harvest of the newly formed shoots. Shoots (each having 2-3 nodes/shoot) thus obtained were also used as a source of nodal explant that gave rise to 1-2 shoots when cultured on modified MS+BAP (0.5 mg/l) medium. Thus, 45-55 shoots could be obtained after 60 days of culture initiation from a single cotyledonary node. About 88% shoots rooted well after 15 hr pulse treatment with IBA (1 mg/l) in liquid MS medium followed by transfer to modified MS medium without IBA. About 80% of these plantlets were successfully acclimatized in plastic pots containing sand and soil mixture and 70% plantlets transferred in the field those survived even after 6 months of transplantation.     

High frequency shoot regeneration from nodal and shoot tip explants in Holarrhena antidysenterica L

Shoot tip and nodal segment explants of Holarrhena antidysenterica when cultured on MS medium containing BAP (1.0-3.0 mg/l) with NAA (0.2-1.0 mg/l) and BAP (1.0-3.0 mg/l) with Kn. (0.2-1.0 mg/l) produced multiple shoots. Maximum multiple shoots was found in MS medium supplemented with BAP (2.0 mg/l) and NAA (0.5 mg/l). Subculture on the same medium resulted in rapid shoot multiplication at an average rate of 16 new shoots per subculture. Addition of urea (100 mg/l) in the medium increased the number of shoots up to 22 per culture. For best rooting, the shoots were excised from the culture flask and implanted individually on half strength MS medium with 0.5 mg/l each of IBA, IAA and NAA. After 20 days of transfer on root induction medium 95% rooting was achieved. Regenerated plantlets were successfully acclimatized and established in soil. About 90% of plantlets survived under open field conditions.     

Rapid in vitro multiplication and ex vitro establishment of Caribbean copper plant (Euphorbia cotinifolia L.): an important medicinal shrub

An efficient micropropagation protocol was developed for E. cotinifolia by utilizing mature nodal segments for axillary shoot proliferation. The nodal explants from a 2-year-old plant were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations (0.5, 2.5, 5.0, 7.5 and 10.0 μM) of 6-benzyladenine (BA), Kinetin and 2-isopentenyl adenine singly as well as in combination with α-naphthalene acetic acid (NAA) or Indole-3-butyric acid (IBA) (0.1, 0.5 and 1.0 μM). The highest regeneration frequency (92 %) with multiple shoots (13.0 ± 1.15) and shoot length (4.23 ± 0.14 cm) was achieved on MS medium supplemented with 5.0 μM BA and 0.1 μM NAA after 8 weeks of culture. Further experiments were performed to test the effects of medium type, medium strength, pH and subculture passages on shoot induction and proliferation. An enhancement in average number of shoots (16.6 ± 0.45) per explant was obtained after four subculture passages. Micro shoots exhibited in vitro rooting on half strength MS medium containing 2.5 μM Indole-3-butyric acid (IBA) after 4 weeks of culture. The in vitro raised healthy plantlets with well-developed roots and shoots were successfully acclimatized in plastic cups containing sterile soilrite for 8 weeks under culture room conditions (150 PPFD) prior to field transfer. Through the acclimatization period (0–56 days), photosynthetic pigments (Chlorophyll a, b and Carotenoid content) decreased during the initial 2 weeks followed by significant increase during the successive period (21–56 days) of acclimatization. At the same time, all the tested antioxidant enzymes (SOD, CAT, APX and GR) exhibited an increasing trend throughout the acclimatization period. The culture room acclimatized plantlets were successfully established in earthen pots containing garden soil in greenhouse with 70 % survival rate.     

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%。     

In vitro photoautotrophic acclimatization,direct transplantation and ex vitro adaptation of rubber tree (<Emphasis Type="Italic">Hevea brasiliensis</Emphasis>)

We investigated the effect of carbon dioxide (CO₂)-ambient (350 µmol CO₂ mol^?1) and CO₂-enriched (1500 µmol CO₂ mol^?1) conditions of in vitro photoautotrophic system on two cultivars, ‘RRIM600’ and ‘RRIT413’ of rubber tree (Hevea brasiliensis) in an acclimatization process of 45 days. Survival percentage of in vitro rubber tree plantlets derived from somatic embryos under ambient CO₂ was better than those under CO₂-enriched conditions, especially in cv. ‘RRIT413’. Subsequently, the survival rate of ex vitro transplanted plantlets was similar to the in vitro plantlets and abnormal morphological characters such as light-green leaves (SPAD), small leaves in cv. ‘RRIT413’ acclimatized under CO₂-enriched conditions were demonstrated 30 days after the plantlets were transferred into the soil. Maximum quantum yield of PSII, photon yield of PSII, stomatal conductance and transpiration rate in cv. ‘RRIT413’ acclimatized under CO₂-enriched conditions were sharply declined by 39.0, 50.6, 47.1 and 45.8%, respectively as compared to those acclimatized under ambient CO₂ conditions. In contrast, the in vitro acclimatized plantlets of cv. ‘RRIM600’ were un-responsive under both ambient- and enriched-CO₂ conditions. In conclusion, genotypic dependent in response to CO₂ enriched conditions in in-vitro acclimatization of rubber tree plantlets was evidently demonstrated as a key result to regulate plant growth and development in ex vitro environments. Interestingly, soluble sugar contents (sucrose, glucose and fructose) were increased after transplanting the plantlets of cv. ‘RRIM600’ acclimatized under CO₂-enriched condition into the soil and thus, can be considered as an adaptive indicator of ex vitro adaptation.