[目的]研究白及在不同郁闭度林下栽培的生长发育特性.[方法]在玉林市六万林场不同郁闭度(0～0.2、>0.2～0.4、>0.4～0.6、>0.6)的八角林林下开展桂及1号白及品种种植试验,分析其不同年龄的生长发育特征.[结果]郁闭度大于0.2时,随着郁闭度增加,白及叶片叶长增长,叶间距加大,但叶片宽度和地径(茎粗)变小;郁闭度大于0.4时,随着郁闭度增加,叶片SPAD值、氮素含量有所降低,而叶片含水量增加;白及生长4年后收获的假鳞茎鲜产量、干物率、干重产量不同郁闭度之间有一定差异,白及品质指标水分及多糖含量无显著差异.在郁闭度大于0.6时,不利于白及产量增加.[结论]品种桂及1号在今后的推广种植中,可在郁闭度小于0.6的林下种植.

[目的]克隆白及磷酸甘露糖变位酶(PMM)基因(BsPMM)cDNA序列,并检测甘露糖合成相关基因的表达特性,为研究甘露糖合成相关基因的调控功能及白及多糖合成机制提供理论依据.[方法]采用RT-PCR克隆BsPMM基因cDNA序列,对其进行生物信息学分析,并采用实时荧光定量PCR(qRT-PCR)检测BsPMM和GDP-甘露糖焦磷酸化酶基因(BsGMP)在不同品种(桂及1号和桂及2号)、生育期(苗期、生长旺期和成熟期)和组织(叶片、茎、假鳞茎和根)中的表达特性,同时测定不同生育期假鳞茎的多糖和甘露糖含量.[结果]克隆获得的BsPMM基因cDNA全长1062 bp,包含一个759 bp的开放阅读框(ORF),编码252个氨基酸,该基因编码的蛋白BsPMM定位于细胞质,含一个从细胞内部到外部的跨膜螺旋区;不稳定系数为41.67,为不稳定蛋白;总平均疏水指数为-0.304,为亲水性蛋白,含植物PMM蛋白特有的4个保守结构域,属于HAD超家族成员.BsPMM蛋白与单子叶植物尤其是兰科植物铁皮石斛和蝴蝶兰PMM蛋白的亲缘关系较近,与罂粟、葡萄和芦笋等双子叶植物PMM的亲缘关系较远.BsPMM和BsGMP基因在不同品种、组织和生育期均有表达,且二者在假鳞茎中表达量显著高于其他组织(P<0.05),区别在于桂及1号在生长旺期的表达量最高,而桂及2号在苗期的表达量最高.桂及1号和桂及2号假鳞茎中甘露糖含量和多糖含量均随生育期推移呈逐渐升高的变化趋势.[结论]BsPMM和BsGMP基因表达具有时空、组织和品种特异性,可能是调控多糖合成代谢途径中的关键基因,参与白及甘露糖和多糖的合成.

黄精在补益类药材市场中是较为常见的药材之一.为了能够满足人们对它的需求,其人工种植成为人们关注的重点和热点.通过对大量资料的搜集和查阅,主要对黄精的栽培技术进行探讨和分析.黄精在繁殖过程中有多种方式,包括种子繁殖、组织培养以及块茎繁殖,大多采取的种植模式是以林下的方式进行套种.种植中主要出现的病虫害类型包括黑斑病、蛴螬等,收获的最佳季节是春季和秋季,最常见的加工方式是蒸煮.

通过抗病品种在病区进行品比和田间抗性评价,测试所选品种的抗性、生育期、产量和主要农艺性状.结果表明:GJ 2和南天黄的抗性表现为高抗,宝岛蕉和中蕉4号为抗,桂蕉1号为高感.GJ 2的生育期为401 d,株产为26.3 kg,农艺性状与桂蕉1号最接近,产量最高,综合性状最优,适宜在广西示范推广.

[目的]克隆白及GDP-甘露糖焦磷酸化酶(GDP-mannose pyrophosphorylase)基因(BsGMP)cDNA全长序列,分析该基因生物学信息,为该基因功能的进一步鉴定提供依据.[方法]基于同源序列克隆GMP基因中间片段,并采用RACE技术扩增GMP基因cDNA全长,用DNAMAN、Tmpred及Softberry等软件进行编码多肽序列、理化性质及亚细胞定位等分析.[结果]从白及叶片中克隆到的GMP基因全长1523 bp,其中包含1086 bp的开放阅读框(ORF),编码361个氨基酸,理论蛋白分子量为39.35 kDa,理论等电点为6.03.进一步分析发现该基因具有跨膜结构,亚细胞定位分析发现该基因主要分布于细胞质和线粒体.蛋白三级结构预测显示,BsGMP蛋白有11个α螺旋,33个B折叠,46个B转角;该蛋白第1～24个氨基酸含有醛酮还原酶的保守结构域,第256～335个氨基酸含有LbetaH家族保守区.BLAST多重序列分析表明,BsGMP氨基酸序列与铁皮石斛、蝴蝶兰的亲缘性最高,分别达到95％和92％.[结论]成功克隆到白及GDP-甘露糖焦磷酸化酶基因(BsGMP),并进行相关生物信息学分析,为该基因的进一步功能研究奠定了基础.

A method for in vitro culture and rapid propagation of Chirita ophio-pogoides was developed using leaves as explants in this study. The results indicat-ed that the medium MS+6-BA 0.1 mg/L+NAA 0.1 mg/L was suitable for bud induc-tion and seedling regeneration from leaves in primary culture. The media MS+0.5 mg/L 6-BA+0.1 mg/L NAA+10% banana+5% potato and MS+0.5 mg/L 6-BA+0.5 mg/L NAA+2% banana were very suitable for cal us multiplication and seedling hardening in subculture, and the proliferation coefficients were 7.9 and 5.6 per 60 d respec-tively. The optimal rooting medium was MS and the rooting rate was 100% on day 30 of culture. The rooted plantlets of C. ophiopogoides were transplanted in green-house with humus soil and 92.5% survived. Theoretical y, using the rapid propaga-tion system, about 20 176 seedlings can be reproduced from a sterile plantlet in a year.

采用组织培养快繁技术培养地枫皮种苗，为人工栽培提供优良地枫皮种苗。以地枫皮的茎段、顶芽作为外植体，用不同的培养基进行诱导培养获得丛生芽，进而生根获得再生植株。结果表明，地枫皮初代诱导较好的培养基为 MS ＋1．5 mg／L 6－BA ＋0．1 mg／L NAA；培养基 MS ＋2．0 mg／L 6－BA ＋1．0 mg／L KT ＋0．3 mg／L NAA 较有利于丛生芽继代增殖，增殖系数为3．2；培养基1／2MS ＋1．0 mg／L IAA 较适宜诱导地枫皮无菌芽的生根；在适宜的基质上移栽，地枫皮种苗成活率为70％。得出的方法具有不受季节和地点限制、繁殖系数较高等优点，随着研究不断深入，技术不断完善，将为地枫皮种苗繁育提供技术支持。

Taking annual Dendrobium officinale as material,the effect of light intensity on the growth and photosynthetic characteristics and the quality of annual Dendrobium officinale were investigated by artificial shading method.The results indicated that the plant height,stem diameter,net presh weight of D.officinale would be the best when the light intensity was at 55%.It had the highest content of polysaccharides when the light intensity was at 43%.

[Objective]To study the system of aseptic seeding and rapid propogationtin of hybrid seeds of Demdrobium officinale and Dendrobium aduncum [Methods] Demdrobium officinale and Dendrobium aduncum were the objects of research,and their hybrid seeds were gained by artificially auxiliary hybridation. The seeds were induced to germinate in vitro,and study on the technology of aseptic seeding was finished. [Results]Successful hybridation of Demdrobium officinale and Dendrobium aduncum could be easily realized,and hybrid fruit-bearing rate could reach 80%; the suitable culture medium for seeds was: MS +1. 0 mg/L 6-BA +0. 1 mg/L NAA,the germination rate after 30 d of cultivation was 89. 7%; appropriate culture medium for proliferation of protocorm was: MS + 1. 5 mg /L 6-BA + 0. 1 mg /L NAA + 100 g /L banana + 1. 0 mg /L AC; appropriate culture medium for differentiation of protocorm was: MS + 1. 5 mg /L 6-BA + 0. 1 mg /L NAA + 200 g /L potato + 1. 0 g /L AC; the formula of culture medium for strenghening the seedlinge and nurturing the rooting was: 1 /2 MS + 0. 6 mg /L NAA +100 g /L banana + 1. 0 g /L AC,the plant grew robustly,the rooting percentage was 100%,and the survival rate of transplanting was higher than 95%. [Conclusions]This method ascertains the optimum culture media formula in the whole regeneration process and all phases from seed germination to acclimatization and transplantation,so fine solidation is laid for its germplasm innovation and industrialized seedling production.

通过HPLC-MS/MS QQQ液相色谱串联质谱联合仪检测分析了铁皮石斛花中的氨基酸组成,并分别采用苯酚硫酸法、NaNO2-Al(NO3)3-NaOH显色体系、Folin-Cioeaheau显色法测定铁皮石斛茎和花的多糖、总黄酮、总酚含量;结合清除DPPH自由基能力、还原力评价铁皮石斛花的抗氧化作用,并与铁皮石斛的茎条进行比较.研究结果表明:铁皮石斛花的氨基酸中主要以精氨酸为主、其次为脯氨酸;铁皮石斛茎条的多糖含量高于铁皮石斛花,而铁皮石斛花的总酚和总黄酮高于铁皮石斛茎;铁皮石斛花的还原力强于铁皮石斛茎.铁皮石斛花有广阔的开发前景.

以10年生南方早熟梨类型“丰水”为试材,在年生长周期中,对春季开花之前、夏季花芽分化期和秋季二次开花期3个不同发育期叶片与芽内源激素水平进行测定并作比较分析.结果表明:南方丰水桑叶片和芽中内源激素含量与生长发育的关系十分密切,芽中低水平的IAA有利于花芽分化,花芽中低水平的IAA有利于花芽春季萌动开放和秋季二次开花的发生;芽和叶片中GA3含量的变化趋势基本一致,高水平的GA3有利于花芽分化;芽中高水平的ZT有利于花芽分化的开始,随着花芽分化进程的推进,芽中的ZT逐渐转移到叶片中;ABA仅在花芽分化期的叶片中出现一次峰值,暗示其在花芽分化中扮演着极其重要的角色.

Tissue culture and rapid propagation of Chirita wentsaii were studied by using the lfeshy leaves as explants. The results showed that the initial medium for the callus induction and seedling regeneration was MS+6-BA 0.1 mg?L-1+NAA 0.1 mg?L-1. The subculture media for bud proliferation and sound seedling cultiva-tion were MS+6-BA 0.5 mg?L-1+NAA 0.2 mg?L-1+10%banana and MS+6-BA 0.5 mg?L-1+NAA 0.5 mg?L-1+10%banana, and their proliferation coefifcients were 6.7 and 4.8 every 60 d respectively. The optimal rooting medium was MS, and the rooting rate was 100%. The rooting plantlets were transplanted in greenhouse and the highest survival rate was about 94%.

以铁皮石斛和钩状石斛为研究对象,通过人工辅助杂交获得杂交种子,诱导种子离体萌发,开展无菌播种技术研究。结果表明,铁皮石斛与钩状石斛杂交容易成功,杂交坐果率可达到80%;适宜的种子萌发培养基为MS+6-BA 1.0 mg/L+NAA 0.1 mg/L,培养30 d时的萌发率为89.7%;培养基MS+6-BA 1.5 mg/L+NAA 0.1 mg/L+香蕉100 g/L+AC 1.0 mg/L对原球茎的增殖最好;培养基MS+6-BA 1.5 mg/L+NAA 0.1mg/L+马铃薯200 g/L+AC 1.0 g/L对原球茎的分化最好;壮苗生根培养基配方为1/2 MS+NAA0.6 mg/L+香蕉100 g/L+AC 1.0 g/L,植株生长健壮,生根率100%,移栽成活率达95%以上。

Bearing trees of Pyrus pyrifolia cv.Hosui in the northern area of Guilin,where physiologically abnormal phenomena were very typical,were selected as the research objects to study the relationship between nutrition of leaves and soil nutrients and its dynamic change rule in different periods by mensurating mineral nutrient contents of leaves and soil in the annual growth cycle,so as to probe into the relationship between the nourishments of leaves and soil in annual growing season. The results were as follows:(1)N,P and K contents of the leaves were abundant in the whole growing season,and the variation range of P contents was the most,while that of N,K contents less,and did not reach a significant level (P0.05). (2)The soil organic matter,hydrolysis of N contents of the pear gardens were very abundant,and effective P and available K contents were generally high. Available N,P,K contents of the soil in the growth cycle reached a significant level(P0.05),and the soil organic matter content was relatively stable. (3)N contents in the leaves had a significant positive correlation with soil organic matter,hydrolysis of N,effective P,and available K content of the soil(P0.05),and P and K contents in the leaves significant negatively and positively correlated with soil hydrolysis of N respectively(P0.05),while had insignificant relationship with soil organic matter,effective P and available K contents of the soil.

Objective To establish the regeneration system of Bletilla striata callus for induction,proliferation,and differentiation.Methods MS was selected as the basic medium.To study the effective factors on induction,proliferation,and differentiation of callus of B.striata with different explants and to optimize the plant growth regulators,kinds,and concentration ratio of plant growth regulators by orthogonal test design.Results The optimal explants were false bulbs of B.striata.MS medium added with 1.0 mg/L 6-benzylaminopurine(6-BA) and 2.0 mg/L 2,4-dichlorophenoxy acetic acid(2,4-D),not only could induce callus formation,but also benefit to the differentiation of callus.MS medium added with 1.0 mg/L 6-BA and 3.0 mg/L 2,4-D promoted the proliferation of callus,and the proliferation coefficient was 7.8.Adding with 0.5 mg/L thidiazuron(TDZ) to the differentiation medium improved the differentiation rate of callus.Conclusion MS + 6-BA 1.0 mg/L + 2,4-D 2.0 mg/L is the optimal medium for the induction of callus;MS + 6-BA 1.0 mg/L + 2,4-D 3.0 mg/L is the optimal medium for the proliferation of callus;MS + 6-BA 1.0 mg/L + 2,4-D 2.0 mg/L + TDZ 0.5 mg/L is the optimal medium for the differentiation of callus.

The explants were obtained from different sections of flower organs of Tacca chantrieri.The effects of different factors on callus induction of Tacca chantrieri,multiplication culture,differentiation of callus,rooting induction and survival rate of culture seeding were investigated using orthogonal design L9(34) and random test.The results showed that the filaments of Tcacca chantrieri were the best explants on callus inducement.The best medium for the first generation callus inducement and the multiplication culture were MS+0.5 mg/L 6-BA +0.2 mg/L KT +2.0 mg/L 2,4-D,relatively better differentiation of callus medium was MS+2.0-3.0 mg/L 6-BA,and relatively better rooting medium was MS+0.3 mg/L NAA.Proper matrix for transplant of Tacca chantrieri culture seedlings were 1/2 fine sand+ 1/2 ash soil.

The technique of the tissue culture and rapid propagation of Euphorbia lathyris were studied. The results showed that the seeds, stems and terminal buds as explants could be induced to sterilized seedings, in which the bud induction rates of stems and the terminal buds were higher. The best medium for multiplication was White+0.05 mg·L-1 TDZ+0.01 mg·L-1 IBA+0.1% AC, and the multiplication coefficient could be 4.5 after 25 d. The best medium for rooting was 1/2MS+1.0 mg.L-1 NAA+0.1% AC. The survival rates reached 60% after transplanting. Vitrification problem in tissue culture of Euphorbia lathyris could be effectively solved by the control of the cytokinin concentration, and materials browning could be alleviated by adding 0.1% AC.

Seeds of Demdrobium officinale could be used as explants to rapid propagate by the way of seed→protocorm→the whole plant→transplant,and medium in each culture stage and some other factors were studied comparatively.The results showed that all seeds of D.officinale which were 60-180 d,especially 150-180 d after pollination could germinate,with a germination rate of 87.2%-94.4%.The suitable medium for seed germination was MS+6-BA 1.0 mg/L+NAA 0.1 mg/L+potato 200 g/L+active carbon 1.0 g/L.The best medium for the formation and multiplication of protocorm was MS+6-BA 1.5 mg/L+NAA 0.1 mg/L+banana mud 100 g/L+active carbon 1.0 g/L,the propagation coefficient was about 20 times/50 d.And the protocorm differentiated on MS+6-BA 1.0 mg/L+NAA 0.1 mg/L+potato 200 g/L+active carbon 1.0 g/L medium,which could have some multiplication in the meantime.After being cultured on MS+6-BA 0.5 mg/L+NAA 0.2 mg/L+banana mud 100 g/L+active carbon 1.0 g/L for a generation,the vigorous plantlets were transferred to rooting medium which was 1/2MS +NAA 0.8 mg/L+mineral salt A 0.2-0.5 mg/L +banana mud 100 g/L+active carbon 1.0 g/L and were cultured for 50-70 d,with 100% rooting rate.Mineral salt A could effectively control the formation of callus or protocorm,and obviously improve the quality and quantity of root regeneration plants.The best time for the transplantation of root regeneration plants was March-May and September-October in Guilin area.The plantlets were transplanted into rotten pine bark handled with high temperature in seedbed of greenhouse resulted in more than 90%survival rates.

Cymbidium floribundum seeds were used to investigate the effects of salt contents,plant growth regulators and light on seed asymbiotic germination.Based on this,the tissue culture and rapid propagation technical system of C.floribundum were established,by investigating the medium formula and culture conditions of proliferation and redifferentiation of protocorm,strong seedlings and rooting of bud.The optimal medium for asymbiotic germination of C.floribundum seeds was 1/6 MS + NAA 0.5mg·L-1+6-BA2.0mg·L-1+meshed potato 50g·L-1+AC1.0g·L-1;the light-intensity of culture was 1.25μmol·m-2·s-1;the seed germination rate was 63.6%.The optimal medium for subculture for protocorm multiplication was 1/4MS+6-BA2.0mg·L-1+NAA0.5mg·L-1+AC1.0g·L-1+PE200g·L-1;the proliferation coefficient was 6.5/60d;the bud redifferentiation rate was 60.2%.The optimal medium for bud redifferentiation was 1/4MS+6BA2.0mg·L-1+NAA0.2mg·L-1+AC1.0g·L-1+PE200g·L-1;the proliferation coefficient was 4.0/60d;the bud redifferentiation rate was 85.0%.The optimal medium for strong seedlings and rooting were 1/6MS+6-BA3.0mg·L-1+NAA1.0mg·L-1+AC1.0 g·L-1+cane sugar 20g·L-1+PE200g·L-1 and 1/4MS+6-BA2.0mg·L-1+NAA1.2mg·L-1+AC1.0g·L-1+cane sugar20g·L-1+PE200g·L-1,respectively.The rooting percentage was100%,and transplantsurvival rate reached 90%.This technology could be used to seedling breeding and germ-plasm protection of C.floribundum.

以茎段和顶芽为外植体,研究不同的基本培养基和植物生长调节剂对鹅不食草组织培养各阶段的影响。结果表明:MS为较适合的基本培养基,初代诱导较好的培养基为MS+6-BA 1.0 mg/L+NAA 0.1 mg/L;继代增殖的最佳培养基为MS+6-BA 1.0 mg/L+NAA 0.3 mg/L,增殖系数为10.4/30 d;生根最佳培养基为:1/2 MS+NAA 0.3 mg/L+IBA 0.1 mg/L。该技术的研究可为鹅不食草的品种改良提供技术平台。