Variation in seed morphometric characters,oil content and azadirachtin content of seeds,in vitro shoot cultures and callus cultures among different populations of Azadirachta indica

We studied seed morphometric characters, oil content and azadirachtin content in a medicinally important and insect repellent tree species Azadirachta indica A.Juss. Ten populations were selected from two central Indian states of Madhya Pradesh and Chhattisgarh and twenty candidate plus trees were selected from each population based on tree attributes. Azadirachtin content was estimated in seeds, in vitro shoots and callus cultures through HPLC. Significant variation(p \ 0.05) was observed in seed morphometric characters, oil content and azadirachtin content among the ten populations. Maximum value for 100 seed weight was obtained in seeds of Raigarh(30.51 g) which was on par with seeds of Bargi(28.37 g)and Chhatarpur(26.36 g). The ratio of highest kernel to seed was also obtained in seeds of Raigarh(60.11%).Kernels from Chhatarpur population recorded the highest oil content(24.20%), which was equal to that in Sihore(23.75%) and Katni(21.75%). We recorded highly significant variation in azadirachtin content in seed samples,in vitro shoot samples and callus cultures. Maximum azadirachtin was present in the seeds of Gwalior(0.882%)which was on par with azadirachtin content in seeds of Chhatarpur, Katni, Sihore, Shahdol and Chhindwara.Maximum azadirachtin in the in vitro shoots was present in the cultures of Gwalior(0.218%) which was statistically on par with Sihore(0.176%). Similarly, in callus cultures of nodal segments maximum azadirachtin was recorded in the Gwalior population(0.033%) which was statistically on par with Sihore(0.028%). The trees of Gwalior and Sihore populations can be produced in large numbers through vegetative propagation to obtain seeds with higher azadirachtin content.     

An efficient in vitro process for recurrent production of cloned plants of Vitex negundo L

An efficient method for cultivation of Vitex negundo L. through axillary shoot (collected from micropropagated plants) proliferation has been successfully developed, which can be employed at a commercial scale. Axillary shoot induction was most successful using nodal explants for propagation on Murashige and Skoog (MS) medium supplemented with various concentrations of single cytokinin or in various combination with auxins. We obtained the maximum percentage (97.6 ± 1.45) response with highest number (16.4 ± 0.60) of shoots per explant on MS medium supplemented with 6-benzyladenine (BA) and ??-naphthalene acetic acid (NAA) at concentrations of 5.0 and 0.5 ??M, respectively. Shoot regeneration frequency was optimized by manipulating pH and using various media. MS medium and pH 5.8 was found to be the optimum for maximum regeneration. Nodal explants from in vitro regenerated microshoots too developed shoots, thus making the process recurrent. Shootlets with 4?C5 nodes were utilized for in vitro rooting, and best response was evaluated on MS medium supplemented with 1.0 ??M indole-3-butyric acid (IBA). The well-developed micropropagated plants were acclimatized (97%) successfully within 2 weeks in soilrite and planted ex vitro in normal garden soil, where they grew well without any morphological and genetic variations. The present regeneration process not only favoured the rapid multiplication but also expressed the regeneration capability of micropropagated plants.     

Variation in seed morphometric characters,oil content and azadirachtin content of seeds,in vitro shoot cultures and callus cultures among different populations of <Emphasis Type="Italic">Azadirachta indica</Emphasis>

We studied seed morphometric characters, oil content and azadirachtin content in a medicinally important and insect repellent tree species Azadirachta indica A. Juss. Ten populations were selected from two central Indian states of Madhya Pradesh and Chhattisgarh and twenty candidate plus trees were selected from each population based on tree attributes. Azadirachtin content was estimated in seeds, in vitro shoots and callus cultures through HPLC. Significant variation (p < 0.05) was observed in seed morphometric characters, oil content and azadirachtin content among the ten populations. Maximum value for 100 seed weight was obtained in seeds of Raigarh (30.51 g) which was on par with seeds of Bargi (28.37 g) and Chhatarpur (26.36 g). The ratio of highest kernel to seed was also obtained in seeds of Raigarh (60.11%). Kernels from Chhatarpur population recorded the highest oil content (24.20%), which was equal to that in Sihore (23.75%) and Katni (21.75%). We recorded highly significant variation in azadirachtin content in seed samples, in vitro shoot samples and callus cultures. Maximum azadirachtin was present in the seeds of Gwalior (0.882%) which was on par with azadirachtin content in seeds of Chhatarpur, Katni, Sihore, Shahdol and Chhindwara. Maximum azadirachtin in the in vitro shoots was present in the cultures of Gwalior (0.218%) which was statistically on par with Sihore (0.176%). Similarly, in callus cultures of nodal segments maximum azadirachtin was recorded in the Gwalior population (0.033%) which was statistically on par with Sihore (0.028%). The trees of Gwalior and Sihore populations can be produced in large numbers through vegetative propagation to obtain seeds with higher azadirachtin content.     

Haploid breeding ofpopulus

After 14 year’s growth observation on the plant growth, it was found that obvious segregation appeared among the haploid pollen plants obtained from anther culture ofPopulus Xiaohei T.S. Hwany ex C. Wang et Tung in vitro. By vegetative propagation, 19 clones were selected and field contrast test was carried out. Statistical analysis result showed a significant difference in volume production. They are superior clones which could be selected from among the pollen plants for utilization. A haploid breeding procedure is suggested in the paper. This study was supported by the National Natural Science Foundation of China     

TLC法测定几种植物中熊果酸的含量

采用薄层扫描法对车前草、夏枯草、山茱萸中熊果酸的含量进行了测定, 其TLC测定条件为用环己烷-氯仿-乙酸乙酯(20 5 8) 作展开剂; 用10%H2SO4乙酸溶液作显色剂。测定结果表明: 3种植物中, 以夏枯草的熊果酸含量最高, 达0 812 %, 车前草的熊果酸含量最低, 为0 516 %, 前者是后者的1 57倍。该方法简便、快速、准确、可靠, 回收率高达96 1 %。     

生长环境及收获期对野鸦椿属植物三萜类化合物含量的影响

【目的】研究不同生境和收获时间对圆齿野鸦椿不同部位总三萜类化合物积累的影响情况,为其药用价值和科学栽培的研究奠定基础。【方法】选择3个低海拔地区的8年生圆齿野鸦椿,分别于2014年11月和2015年3月采集其叶片、枝条和果实,选择高海拔地区6年生的野鸦椿和圆齿野鸦椿,于2014年8月分别采其叶片和枝条,对其总三萜类化合物含量进行了测定和比较分析。【结果】1)8月采集的叶片中三萜类化合物的含量,野鸦椿高于圆齿野鸦椿,且其含量差异呈显著性水平;但是,8月采集的枝条中三萜类化合物含量,野鸦椿低于圆齿野鸦椿。2)圆齿野鸦椿不同部位的总三萜类化合物含量存在差异,且2个月份采集的含量高低顺序均为叶片>枝条>果皮>种子,均以叶片中的含量为最高,依次为9.06%(农大)、7.14%(清流)和6.85%(邵武)。3)11月采集的圆齿野鸦椿样品中的三萜类化合物含量高于3月所采样品中的含量,其叶片中的总三萜类化合物含量最多可提高172.91%。4)圆齿野鸦椿叶片、果皮和种子中的三萜类化合物含量与试验地的海拔高度、年降水量均表现出极显著的负相关关系,与年均气温存在极显著的正相关关系,其叶片中的三萜类化合物含量还与年均无霜期呈正相关关系,其枝条中的三萜类化合物含量与各地理和气象因素之间的相关性均不显著。【结论】圆齿野鸦椿的叶片采收期长于野鸦椿;低海拔、较少降水量和年均气温较高有利于圆齿野鸦椿植株对三萜类化合物的累积,这3个因子可以作为圆齿野鸦椿适宜栽培区域的划分指标;夏季采摘野鸦椿叶片,秋冬季采摘圆齿野鸦椿叶片,所采叶片的总三萜类化合物的含量均较高。     

Improvements in asexual multiplication procedures for jack pine (Pinus banksiana)

Various pre- and post- harvest factors wereinvestigated to refine asexual propagation proceduresfor jack pine (Pinus banksiana) usingproliferated dwarf shoot (PDS) cuttings. The factorsinvestigated included donor plant cultural treatments(growth conditions, fertility, pruning and coldtreatments) and application of auxins to cuttings topromote rooting. Production of PDS under greenhouseconditions was enhanced by pruning, fertilizerapplications and a cold treatment. Coldstorage of donor plants (2–6 weeks at 2°C)prior to cutting harvest, increased rootingfrequencies. After a 9-month seedling greenhousegrowing phase between 13 and 34 PDS cuttings could beharvested from a donor plant. Three pruningtreatments followed by successive harvests of the PDSdeveloping on the most distal 2 cm of the shootsresulted in the highest PDS yields. Optimal rootingwas obtained from donor plants grown in a greenhouseenvironment with relatively low fertilizerapplications (150 ppm N; 20-8-20 monthly). Rooting ofPDS cuttings ranged from 45–100%. Rootingfrequencies were highest with 5.4 mM NAA application. Implications of these results with respect tooperational asexual propagation of jack pine arediscussed.     

山杏叶桦木酸、齐墩果酸和熊果酸的研究

我国山杏资源十分丰富,是绿化荒山的优良树种。采用HydersirBDSC18色谱柱（150×4．6mm,5μm）,流动相体积比为90：10的0．3‰硫酸甲醇溶液一3‰硫酸水溶液,检测波长210nm,建立了山杏叶中三萜酸测定的高效液相色谱法。结果表明：桦木酸、齐墩果酸与熊果酸的浓度在30～2000μg／mL（r桦=0．9971,r齐=0．9986,r熊=0．9978）范围内与色谱峰面积呈良好线性关系;平均加标回收率分别为99．9％、101．1％和98．9％。以山杏叶为材料,采用高效液相色谱法和紫外可见分光光度法分别对山杏叶中的三萜酸成分进行分析,结果表明,山杏叶富含三萜酸,落叶期山杏叶中总三萜酸（17078#g／g）、桦木酸（1842μg／g）、齐墩果酸（2536μg／g）和熊果酸（5706μg／g）含量均高于非落叶期。本研究首次建立了山杏叶中桦木酸、齐墩果酸和熊果酸同时测定的反向高效液相色谱法（HPLC）,发现落叶期山杏叶富含三萜酸,为山杏叶综合开发利用提供了基础数据。     

Inhibition of Trypanosoma cruzi by plant extracts used in Chinese medicine

In this work, we assessed the effect of extracts obtained from 17 plants used in traditional Chinese medicine. These extracts were tested in vitro with the epimastigote form of Trypanosoma cruzi, clone Bra C(15) C(2), at 27 degrees C in F-29 medium at a concentration of 100 microg/ml in axenic cultures. Allopurinol was used as reference drug. Seven plant extracts showed inhibitory activities lower than 25%. Pueraria lobata, Mahonia beaei, Dictamus dasycarpus, Kochia scoparia, Sophora flavescens and Ligustrum lucidum showed effects with inhibition values between 25% and 60%, whereas Lithospermum erythrorhizon, Saussurea lappa, Melia toosendan and Cinnamomum cassia showed the greatest inhibitory activity of 100%. The IC(50) of these extracts were: 0.4, 2.4, 1.8 and 3.9 microg/ml, respectively. The MTT assay was made and did not show cytotoxic activity. These results allowed us to suggest that L. erythrorhizon, S. lappa, M. toosendan and C. cassia could be a source of new compounds against T. cruzi.     

越南多毛金花茶的光合特性和叶解剖结构对光照环境的适应

[目的]以越南多毛金花茶(Camellia hirsuta Hakoda et Ninh)为实验材料,开展不同光照处理对多毛金花茶光合特性、叶解剖结构及光合色素含量的影响研究,为越南金花茶引种栽培和抚育管理提供理论依据。[方法]于2017年5月上旬,将越南多毛金花茶置于不同光照强度(10%、20%、30%、40%、50%,通过覆盖不同层数的遮荫网获得)的遮荫棚内生长,处理时间2个月。[结果]不同光照处理下,多毛金花茶净光合速率日变化趋势不同,20%、30%光照处理显著提高了多毛金花茶叶片净光合速率、蒸腾速率,同时蒸汽压亏缺降低,促进光合作用的进行,达到显著差异(P0.05);在30%光照处理下,光合色素的含量最高,而叶绿素a/b的值最低,30%光照更有利于多毛金花茶叶片中叶绿素b的合成。20%、30%光照处理下的多毛金花茶叶片具有更发达的栅栏组织、海绵组织及更厚的叶片厚度,这2个处理的叶片结构可能有利于其对环境的适应并且光合能力更强。在适当的遮荫条件20%～40%光照处理下,多毛金花茶的最大净光合速率增大。[结论]在20%、30%光照处理下,其叶片具有更发达的叶片厚度和栅栏组织,气孔密度相对较低,净光合速率、蒸腾速率得到提高,胞间CO_2浓度和蒸汽压亏缺降低,CO_2和水蒸气进出气孔阻力变小,同时在30%光照处理下光合色素的含量最高,说明光合速率得到有效提高。越南多毛金花茶适宜在20%～30%光照强度的环境下生长。     

东北林区森林凋落物中的有机酸分析

落叶松(Larixolgensis)、水曲柳(Fraxinusmandshurica)、红松(Pinuskoraiensis)、白桦(Betulaplatyphylla)是东北林区主要造林树种。于2002年10月初,在东北林业大学帽儿山实验林场老爷岭和尖砬沟试验站(45?2?45?0碞,127?0?127?8碋)采集了4个树种林分的森林凋落物样品。采用气相色谱法对当年新鲜凋落物及半分解凋落物中有机酸进行了定性、定量分析。鉴定出13种有机酸,包括草酸、丙二酸、反丁烯二酸、丁二酸、顺丁烯二酸、苹果酸、柠檬酸、棕榈酸、硬脂酸、油酸、亚油酸、亚麻酸、花生酸。结果表明:对于新鲜凋落物而言,所测的7种低分子量有机酸中草酸含量最大,一般高于30mg/g;6种高级脂肪族酸中油酸或亚油酸含量最大,高于40mg/g;在半分解凋落物中,草酸含量最多,其次为亚油酸,再次为油酸;同一树种新鲜凋落物中有机酸种类和含量明显多于半分解凋落物。图5表2参10。     

安徽地区华东野核桃果实特征和坚果品质的初步评价

[目的]探究华东野核桃果实性状、核仁成分及其相关性,为其资源的开发和综合利用提供依据。[方法]本研究以皖南华东野核桃资源为对象,测定其鲜果单果质量、果皮厚度与坚果单果质量、壳厚度、出仁率等果实性状及坚果核仁成分,分析其相关性。[结果]华东野核桃果实性状具有丰富的变异,大部分变异系数大于10%,其中,坚果壳厚度变异系数最大为43.92%。对坚果核仁成分的测定表明:核仁中脂肪含量最高,蛋白质次之,总糖含量较低;对脂肪组分的进一步分析可知,其不饱和脂肪酸含量较高,达92.68%。利用相关性分析,发现坚果单果质量与核仁脂肪含量呈显著正相关。利用主成分分析对华东野核桃果实性状及核仁成分的综合分析及评价发现,单株编号10和8的鲜果大小及不饱和脂肪酸含量在所受试单株中综合表现最好,可作为进一步育种的遗传资源。[结论]本研究首次报道华东野核桃果实核仁成分,发现其核仁脂肪酸含量高于薄壳山核桃、山核桃,且不饱和脂肪酸含量也高,表明其核仁是值得开发的一种新食品资源。该研究结果为华东野核桃资源的综合开发与利用提供了数据支持。     

Rapid in vitro cloning of a 40-year-old tree of <Emphasis Type="Italic">Azadirachta indica</Emphasis> A. Juss. (Neem) employing nodal stem segments

An efficient in vitro process for rapid clonal propagation of a 40-year-old tree of Azadirachta indica employing nodal stem segments was developed. Season of collection and maturity of explants showed direct influence on bud-break. Nodal stem segments collected during the month of April gave best response. Maximum bud-break (78.6–81%) was obtained when middle order nodes (3rd or 4th node from apex) were taken. Amongst different cytokinins used, 6-benzylaminopurine (BAP) at the concentration of 1.11 μM was found most effective in inducing multiple shoots, whereas inorganic and organic constituents of the medium influenced growth and general condition of proliferating shoots. On an average 3.1 shoots per explant were regenerated in modified Murashige and Skoog medium supplemented with 1.11 μM BAP, 1.43 μM indole-3-acetic acid (IAA) and 81.43 μM adenine hemisulphate. Isolated shoots were rooted in presence of 2.46 μM indole-3-butryic acid (IBA). Root induction took place in 8–10 days with 100% rooting. The in vitro-raised plantlets were successfully transplanted in potted soil and finally grown under field conditions with 100% survival. The genetic fidelity of such in vitro-raised field-grown plants was ascertained by random amplified polymorphic DNA (RAPD) markers. Furthermore, the azadirachtin content of in vitro-cloned plants was found comparable to the mother tree proving their chemical stability also. The protocol developed holds good for in vitro cloning of mature elite neem trees.     

Triterpenoid content and expression of triterpenoid biosynthetic genes in birch(Betula platyphylla Suk) treated with 5-azacytidine

DNA methylation is widespread in plants and associated with plant development and defense mechanisms. However, the relationship between DNA methylation and plant secondary metabolism has rarely been reported. Here, when birch suspension cells were treated with 5-azacytidine(5-azaC), which blocks DNA methylation, triterpenoid accumulation was significantly promoted and antioxidant and defense enzymatic activity changed.For studying triterpenoid accumulation, 0.1 m M aza C was optimal. A qRT-PCR assay revealed increased expression of genes encoding key triterpenoid biosynthetic enzymes.Evaluation of methylation polymorphisms at CCGG sites showed that the methylation level was lower in cells treated with 5-azaC. These results demonstrated that 5-aza C treatment led to an increase in the production of triterpenoids in cell cultures through a mechanism that involved in DNA methylation, which resulted in the induction of genes encoding the key enzymes. The study providesevidence of a relationship between DNA methylation and regulation of secondary metabolism.     

LED光质对油茶苗生长和光合特性的影响

[目的]探寻适合油茶苗培育的最佳LED光质配比。[方法]以2年生长林4号油茶扦插苗为试材,LED白光为对照,研究LED红光、蓝光及其复合光对油茶苗生长和光合特性的影响。[结果]表明:L1(红光)处理下,油茶苗高生长量最大,但表观量子效率、光补偿点和胞间CO2浓度最小;L7(蓝光)处理下,油茶苗光补偿点最高,但高生长量、干物质含量、壮苗指数、气孔导度和蒸腾速率最小;L6(10%红光+90%蓝光)处理下,油茶苗地径生长量、干物质含量、壮苗指数、光合色素含量、表观量子效率、最大净光合速率、暗呼吸速率、气孔导度、蒸腾速率、实际光能转换效率和相对电子传递速率均最大;L4(50%红光+50%蓝光)处理下,油茶苗地径生长量、光合色素含量、最大净光合速率、实际光化学效率和相对电子传递速率均最小,且非光化学淬灭系数最大。[结论]与对照及其它处理相比,L6(10%红光+90%蓝光)处理更利于长林4号油茶苗生长和光合能力的提高,是培育油茶壮苗较为理想的光质。     

孟加拉国Madhupur林区内不同森林树种中丛枝菌根多样性的研究

从 Madhupur 林区的不同地点采集了 Acacia auriculi-formis A. Cunn. ex Benth., A. mangium Wild., Artocarpus het-erophyllus Lamk. C., Dalbergia sissoo Roxb. ex A. P. D., Euca-lyptus camaldulensis Dehnn., Hevea brasiliensis (Wild. ex Juss) Muell. Arg., Swietenia macrophylla King. and Tectona grandis L. 8个树种的根和根际土。采用简单常规的根围土壤真菌孢子分离、形态鉴定和树木细根染色、显微镜观察等方法,进行了孟加拉国Madhupur林区内不同森林树种中丛枝菌根多样性的研究。图2表2参46。     

Direct shoot organogenesis from leaf explants of Embelia ribes Burm. f.: a vulnerable medicinal plant

An efficient system was developed for direct plant regeneration from in vitro-derived leaf explants of Embelia ribes Burm. f., a vulnerable medicinal woody climber of the Western Ghats of India. The in vitro procedure involved three steps that included induction of shoot initials from leaf tissue, regeneration and elongation of shoots from the shoot initials, and rooting of shoots. The induction of shoot initials was achieved on Murashige and Skoog (MS) solid medium supplemented with different concentrations of thidiazuron (TDZ). The best medium for shoot induction was MS with 0.272 μM TDZ. Numerous shoot primordia developed within 2–3 weeks on the leaf margin as well as on the midrib region, without any callus phase. In the second step, the shoot clumps separated from the leaf explant on transfer to MS basal medium, resulting in the differentiation of 90% of the shoot initials into well-developed shoots. The 2- to 3-cm-long shoots rooted on half-strength MS basal medium supplemented with 4.90 μM indole-3-butyric acid (IBA) and 3% (w/v) sucrose in the third stage. The rooted plants could be established in soil with 70% success. This protocol could be utilized for in vitro propagation and conservation of this important threatened medicinal plant.     

Reproductive capacity of first-generation adults of the rice water weevil<Emphasis Type="Italic"> Lissorhoptrus oryzophilus</Emphasis> Kuschel (Coleoptera: Curculionidae) in Zhejiang,China

First-generation adults of the rice water weevil, Lissorhoptrus oryzophilus Kuschel were collected from Yueqing (28.1°N latitude, 120.9°E longitude), Zhejiang, China, during late June and mid-July and reared on rice plants (cultivar Shanyou 63) in the laboratory (28±1 °C, 16L:8D) to examine their reproductive capacity. On plants aged 16–22 days, adults from the field and collected in mid-July could reach oviposition onset more rapidly and deposit more eggs than those collected earlier. Adults that emerged in late June and early July displayed a greater tendency to become reproductive, compared with those which emerged in mid-July. However, each of these groups of adults, either field-collected or newly emerged, deposited only 3–14 eggs within 60 days of feeding. Reproductive capacity was greatly related to plant age, with plants aged below 20 days being most favorable; on plants aged over 20 days, reproductive development could take place, but little oviposition was observed. These results indicate that the reproductive capacity of first-generation adults in Zhejiang is dependent upon the time of adult emergence and post-emergence, as well as on the age of rice plants.     

Vegetative propagation of Colutea arborescens L., a multipurpose leguminous shrub of semiarid climates

The vegetative propagation of the multipurpose leguminous shrub Colutea arborescens from leafless hardwood cuttings was examined. Specific attention was paid to the factors: 1) cutting origin (apical, medial or basal); 2) indole-3-butyric acid (IBA) treatment (0, 50 or 200 ppm); and 3) relative humidity of the propagation environment (high or low humidity). After 120 days, 67.6% of the C. arborescens cuttings had rooted. The rooting ability of the cuttings was significantly influenced by the cutting origin and hormonal treatment. No effects on rooting percentage were shown by the relative humidity of the propagation environment. The best rooting response (95%) was shown by basal cuttings treated with 200 ppm IBA. Cutting origin, IBA treatment and relative humidity were significantly related to the root number, longest root length and the fresh/dry biomass of roots and leaves. Under conditions of high humidity, basal cuttings treated with 200 ppm IBA gave rise to the most highly developed plants. The positive relationship between sprouting and rooting suggests that successful propagation is related to the shoot formation potential of the cutting's food reserves. These findings were used to propose an efficient method for the vegetative propagation of C. arborescens plants. This revised version was published online in June 2006 with corrections to the Cover Date.     

Factors influencing in vivo and in vitro micrografting of sandalwood (Santalum album L.): an endangered tree species

In vivo micrografting of Santalum album was achieved (50%) by grafting 4- to 5-cm-long scions, collected from a candidate plus tree (CPT) of 50–60 years of age, onto 90-day-old nursery-grown rootstock. Scion size, rootstock age, and scion collection season were found to influence graft success. Grafted plants were incubated under greenhouse conditions for 6–8 weeks during the graft union process. In vitro micrografts were achieved by placing 1- to 2-cm-long scions derived from nodal shoot segments (collected from CPT) onto the hypocotyl of 45-day-old in vitro rootstocks. Use of in vitro grown shoots as a source of scion gave better graft success (60%) than scions collected directly from field-grown trees. In vitro grafting was also influenced by scion size and rootstock age. Under favorable conditions, scions and hypocotyls unite to form complete plants that produced two to four leaves after 6–8 weeks. This is the first report on in vivo and in vitro micrografting of S. album having potential for production of disease-free clonal plants for conservation and improvement targets.