Experimental Research on Four Season Soilless Cultivation of the Sprouting Seedlings of Toona sinensis in Chongqing

Using seed sowing cultivation method, the four-season soilless cultivation of the sprouting seedlings of Toona sinensis was carried out with different germination accelerating treatments, cultivation substrates, cultivation environment and management technologies by using the corresponding treatment methods in different seasons, so as to produce the sprouting vegetable products of T. sinensis which was not restricted by seasons. The test results showed that using the mixed substrate of pedite and vermiculite in winter and spring can keep moisture but not too high, and the produced sprout vegetable had good quality and high yield.

Design and Construction of Rainwater Harvesting and Water-saving Irrigation System of Toona sinensis on Mountain Slopes

In order to collect rainwater and resist drought to enhance the utilization rate of rainfall and water resources, through project rainwater harvesting measures, the total annual rainwater harvesting amount of the six greenhouses was calculated according to annual average precipitation 542.2 mm, up to 1 095.7 m^3. The upper natural slopes of cultivated land were as rainwater harvesting areas, and total annual rainwater harvesting amount was 49 242 m^3 on the mountain slopes with an area of 73.37 hm^2, while total water storage amount was 39 394 m^3 in theory, so it could meet water use for the irrigation of 26.28 hm^2 of T. sinensis land. To be convenient for rainwater harvesting, irrigation and supplying water to the water-saving cellars, one pert-cut and part-fill reservoir （which was 470 m^3 in volume） was built on the mountain slopes at the right rear of the greenhouses, and their altitude difference was 50 m. The reservoir was sealed and was built with reinforced concrete. Water-saving cellars were distributed in front and the middle and at the back of two rows of greenhouses, and they were connected with each other. The reservoir could supply water to the water-saving cellars and also collect water by mountain slopes, from the lower water-saving cellars or deep wells. Two rainwater hervesting ditches that were 1 650 m in length were at the lower edge of arable land in the upper reaches of slopes to intercept rainfall runoff and make it flow into channels and then the sedimentation tanks. The total annual rainwater harvesting amount of the reservoir and water-saving cellars was 1 222.5 m^3.

Bioactivities of Extracts from Toona sinensis against Sitophilus zeamais

[ Objective ] The repellent activities and contact toxicity of extracts from Toona sinensis against Sitophilus zeamais were studied. [ Meth- od] 95% alcohol, petroleum ether and dichloromethane were selected to extract the active components from the leaves and barks of T. sinensis, respectively. And then the extracts were prepared at 0.6, 0.3, 0.15 g/ml to study the biological activity against S. zeamais in lab. [ Result J The petroleum ether extracts from the leaves of T. sinensis had significantly repellent effects on S. zeamais, but the extracts from T. sinensis with three solvents had no obvious contact toxicity against S. zeamais. [ Conclusion ] The petroleum ether extracts from S. zeamais had certain repellent activity.

香椿(Toona sinensis)微卫星富集文库构建及特性分析

采用Dynal磁珠富集法构建了香椿微卫星富集文库,通过测序结果对文库的特性进行了分析。实验使用改良CTAB法提取香椿基因组DNA,用（AC）8、（AG）8和（ATG）123种带有生物素标记的探针与香椿基因组DNA的酶切片段进行杂交,将磁珠捕获的含有微卫星序列的DNA片段插入p MD 18-T载体,并转入感受态细胞Trans 5α构建克隆,经筛选后得到含356个克隆的香椿基因组微卫星富集文库。从富集文库中挑选插入片段长度为400-800 bp的128个克隆进行测序,其中含有SSR的序列77条,得率达60.16%,其中完美型占82.69%,非完美型8.65%,混合型8.65%。上述结果为SSR位点的进一步开发奠定了基础。

Analysis of Volatile Compounds and Identification of Characteristic Aroma Components of <i>Toona sinensis</i>(A. Juss.) Roem. Using GC-MS and GC-O

In this study, volatile compounds present in Toona sinensis (A. Juss.) Roem (TS) were investigated and their characteristic aromatic components were identified using Headspace Solid-phase Microextraction (HS-SPME) followed by Gas Chromatography-Mass Spectrometry (GC-MS) and Gas Chromatography-Olfactometry (GC-O). The optimum conditions for extracting the volatiles from TS were achieved with the experimental parameters including the use of a 65 μm polydimethylsiloxane/divinyl benzene (PDMS/DVB) fibre, an extraction temperature of 40℃ and an extraction time of 30 min. Under these conditions, 56 volatile compounds were separated and 53 were identified by GC-MS. Among them, 21 sulfide compounds (42.146%) and 27 terpenes(55.984%) were found to be the major components. The sample was analyzed by GC-O and 26 elutes were sniffed and their sensory descriptions evaluated by an odor panelists. Analysis of the data indicated, two compounds cis and trans isomers of 2-Mercapto-3,4-dimethyl-2,3-dihydrothiophene were major contributors to the characteristic aroma of TS.

Lactam Triterpenoids from the Bark of Toona sinensis

Three new limonoid-type triterpenoids,namely toonasins A–C(1–3)with a rare lactam E ring,along with six known compounds(4–9)were isolated from the barks of Toona sinensis.The structures of new compounds were elucidated by interpretation of spectroscopic data,and the relative configuration of compound 1 was further characterized by X-ray crystallographic analyses.The isolated compounds were evaluated for their cytotoxic activities against five human tumor cell lines(HL-60,SMMC-7721,A-549,MCF-7 and SW480),and compounds 3 and 5 showed weak cytotoxicities.

Experimental Study on Cultivation of Multi-head Seedlings of Toona sinensis in Chongqing

Seedlings of Toona sinensis were planted in seeding and breeding ways to increase the yield of T. sinensis by raising the number of lateral branches of T. sinensis seedlings by means of cutting and pinching through sowing and breed- ing in containers and sand bed nursery. The results showed that： after cutting and pinching, seedlings with multi-lateral branches could be formed. The cutting treat- ment in the container had a small number of seedlings with over 2 lateral buds, and the lateral buds were weak. Some of the weak lateral buds disappeared with the growth going on, showing poor growth performance. The pinching treatment in the sand bed nursery found a large number of seedlings with more than 2 lateral buds, and the lateral buds could grow into lateral branches, forming the T. sinensis seedlings with multi-lateral branches. Proceed to cutting processing, the secondary and third-level lateral branches came into being, which could make the seedlings of T. sinensis have multi-head tree shape in 2-3 years.

Research of Protected Cultivation Techniques of Toona sinensis on Mountain Slopes

In order to prolong the supply period and increase the economic value of Toona sinensis,the protected cultivation of T. sinensis was carried out on mountainous slope to achieve stagger-season planting and the year round supply. The results showed that in late October to midNovember,the temperature was moderate( 1-10℃) and suitable for the lifting of seedlings and temporary planting dormancy. The line spacing of rooted seedlings in greenhouse was( 22-24) cm ×( 4-5) cm and planting density was 900 000-1 200 000 seedlings/ha. The line spacing of seedlings was( 14-13) cm ×( 3-4) cm and planting density was 1 950 000-2 100 000 seedlings/ha. T. sinensis began to sprout at 8-10℃ and stretch at 10-12℃. When the temperature was stable at 14℃ or above,T. sinensis started bolting and emerging leaves. The terminal buds sprouted in 40-50 d for the T. sinensis planted in greenhouse with the day temperature of 15-22℃ and night temperature over 10℃; the terminal buds sprouted in about 15 d for those planted in the greenhouse with day temperature of 20-25℃ and night temperature over 10℃. T. sinensis could be picked 3 times when planted in greenhouse,and the yield was about 24 720 kg/ha.

A likely autotetraploidization event shaped the Chinese mahogany(Toona sinensis)genome

Chinese mahogany(Toona sinensis) is of considerable medical and economic importance, and its genome has been deciphered. However, the process underlying its polyploidy is unclear, and the chromosomal evolutionary trajectory is poorly understood. Here, by reanalysing the T.sinensis genome, we found evidence of a tetraploidization event(T. sinensis special tetraploidization, TST) that occurred approximately 15-17 million years ago(MYA) after the core eudicot-common hexaploidization(ECH or gamma) event. We characterized the synonymous nucleotide substitution rates(Ks values) of collinear genes and found that T. sinensis genes affected by the TST evolve at a slower rate than Acer yangbiense genes. Furthermore, we identified homologous genes related to polyploidization and speciation and constructed multiple alignments with different reference genomes. Notably, the significant balance of gene retention and loss characterized in the two TST-derived subgenomes suggests an autopolyploid nature of the TST. Moreover, we deduced the chromosomal karyotypes of the two subgenomes and identified 7chromosomal fusions that have shaped the T. sinensis genome;more information is available on a newly constructed karyotype platform(http://www.cgrpoee.top/Toona_sinensis/index.html). The T. sinensis genome preserves the ancestral chromosome structure of dicotyledons well and could serve as a good reference for understanding genomic changes in other Meliaceae and Sapindales plants. In addition, we verified that tandem duplication and the ECH have promoted the expansion of terpene synthase(TPS) genes;conversely, the TST seems to have inhibited expansion of these genes. This present effort has clarified the polyploidy events of the T. sinensis genome, filled gaps in the history of karyotype evolution, and laid a solid foundation for further genomic studies in the Meliaceae research community and beyond.

Study on the relationship between Toona sinensis Roem stand productivity and site conditions in Sichuan Basin

According to the relationship between Toona sinensis Roem stand volume,productivity and forest age,site conditions,stand density and other factors,through selecting 8 representative cities or counties,using standard investigation and stem analysis method,this paper makes relatively systematic research about the Toona sinensis Roem plantation accumulation,productivity,and the relationship between them and site conditions.Through comparative analysis indicators of multiple site conditions,this paper expects to give a more comprehensive picture about the source of accumulation and productivity difference.

Effects of Drying and Blanching on the Quality of Toona sinensis

[Objective]To explore the best T.sinensis processing technology.[Method]The T.sinensis was treated by hot wind drying,freeze drying and blanching,and the contents of ascorbic acid,total phenolics and nitrite were determined respectively before and after the treatment to be a standard to evaluate the influence of different processing methods on the quality of T.sinensis.[Result]Both drying and blanching produced certain impact on the contents of all three components,among which the content of ascorbic acid was slightly influenced by vacuum freeze drying and short-time blanching,but greatly affected by long-time blanching and hot wind drying;the loss rate of polyphenols was above 60% when the hot wind drying,freeze drying and blanching lasted for more than 5 min,while it experienced little change under short-time blanching;hot wind drying and blanching may greatly reduce the content of nitrite,however,the vacuum freeze drying even improved the content,which would produce adverse impact on food safety.[Conclusion]Short-time blanching is the best method to process T.sinensis,it can effectively reduce its content of nitrite and retain the nutrients.

Effect of Low Temperature Stress on Protein Content of Toona Sinensis Seedlings and Expression of Specific Protein

[Objective] The paper was to study the changes of protein content and component in leaves of Toona Sinensis under low-temperature stress.[Method] Potted experiment was adopted,and SDS-PAGE electrophoresis was also used to analyze the dynamic changes of protein in T.Sinensis leaves.[Result] Low temperature stress could change the content and component in leaves of T.Sinensis,but different provenances had different performance under different temperatures,so did the same provenance under different stress periods.SDS-PAGE electrophoresis results indicated that the change law of provenances of Xixia of Henan and Nanjing of Jiangsu with stronger cold resistance was similar,showing the change trend of "increase-decrease-increase".Protein was greatly expressed after stress for 1 d,the color of band became darker;the content gradually decreased in the following second and third day,and the color of bands was lighter;the content began to increase at the forth day.Two provenances induced the specific proteins with molecular weights of 27.6 and 22.5 kD,respectively.The soluble protein content of provenance of Xiapu of Fujian with relatively weak cold resistance was gradually increased,but no new protein bands were induced.The changes of protein band color of various provenances in SDS-PAGE electrophoresis was basically consistent with the changes of protein content,the provenance with stronger cold resistance could induce the production of specific proteins.[Conclusion] The study provided theoretical basis for under the molecular mechanism of plant cold resistance,which had great significance in theory and practice.

Four New Limonoids from the Barks of Toona ciliata

Four new limonoids,toonayunnanaes F−I(1−4),and six known compounds(5−10)were isolated from the barks of Toona ciliata.Their structures were elucidated by thoroughly analyzing of NMR and HRMS data,and single-crystal X-ray diffraction of 1.The oxetane ring moiety in 1 was rare in limonoids and other natural products.Compound 1 showed nitric oxide(NO)inhibitory effect with an IC5038.45±0.41μM in lipopolysaccharide(LPS)-activated RAW 264.7 macrophages.

基于GC-IMS和GC-MS技术结合化学计量法分析干燥方式对香椿挥发性成分的影响

为研究干燥方式对香椿挥发性风味物质的影响,利用气相色谱-离子迁移谱技术(gaschromatography-ion mobility spectrometry,GC-IMS)和气相色谱-质谱联用技术(gas chromatography-mass spectrometry,GC-MS)、相对香气活性值(relative odor activity value,ROAV)对真空冷冻干燥、热泵干燥、热风干燥、微波真空干燥的香椿样品进行挥发性成分分析,并进一步结合化学计量方法主成分分析(principal component analysis,PCA)和偏最小二乘判别分析(partial least squares-discriminant analysis,PLS-DA),探究样品间挥发性成分的差异性。结果表明,通过GC-IMS检测到78个峰,鉴定出59种化合物。通过GC-MS鉴定出94种挥发性化合物,主要挥发性成分是硫化物、烯烃类和醛类。经过干燥后总挥发性化合物的含量显著降低。通过ROAV法确定了(E,Z)-二-1-丙烯基二硫化物、2-巯基-3,4-二甲基-2,3-二氢噻吩、正己醛、2-己烯醛、丁香酚等为香椿样品中关键香气化合物。PCA和PLS-DA结果表明,不同干燥方式的样品与新鲜样品有明显的分离,彼此之间也有明显的区别。鲜样和真空冷冻干燥样品为一组,其他3个干燥香椿样品为一组。根据变量投影重要性(variable importance in projection,VIP)得分共筛选出10种(VIP>1)标志挥发性化合物。干燥方式对香椿风味特性影响显著,真空冷冻干燥组与鲜样最为接近,考虑实际应用热泵干燥为香椿最适宜的脱水方式。本研究为热加工过程中风味品质控制提供一定的理论参考。

浙江省香椿溃疡病病原菌的分离和鉴定

[目的]分离和鉴定引起浙江省香椿溃疡病的病原菌,揭示香椿溃疡病的病因,为该病害的科学防控提供理论依据。[方法]观察香椿溃疡病的田间症状,使用常规组织分离法分离病原菌,利用形态学结合分子生物学方法对病原菌进行物种鉴定,采用柯赫氏法则验证分离菌株的致病性。[结果]1)从感病香椿样品中分离到2株病原菌;病原菌的培养性状、有性世代和无性世代的形态特征与假毛丛赤壳菌一致。2) LSU rDNA、ITS rDNA和tef1-α基因序列分析以及多基因系统发育分析表明,XCKY2菌株的LSU rDNA、ITS rDNA和tef1-α基因片段与假毛丛赤壳菌的同源性均为100.00%;XCKY1和XCKY2菌株与11株假毛丛赤壳菌聚为一支,贝叶斯后验概率为0.94。3)致病性测定结果表明,假毛丛赤壳菌的菌饼和分生孢子悬浮液均能引起健康香椿苗发病,从发病香椿苗组织中也能再分离到假毛丛赤壳菌,由此确定假毛丛赤壳菌为香椿溃疡病的病原菌。[结论]从浙江省感病香椿样品上分离到2株假毛丛赤壳菌,经鉴定为引起香椿溃疡病的病原菌。本研究首次报道假毛丛赤壳菌是香椿溃疡病病原菌。

香椿老叶化学成分及生物活性研究进展

香椿老叶资源丰富,且其中含有大量具有生物活性的化学成分。为了对香椿老叶进行合理的资源开发利用,推动香椿产业的发展继而推动乡村振兴。本课题组将近年来各研究组对香椿老叶的化学成分和生物活性的研究进行了系统的梳理,发现香椿老叶中含有大量的黄酮类、多酚类、皂苷类、多糖类等化学成分,具有显著的抗菌、抗氧化、提高动物生长性能等生物活性。并对香椿老叶的后续研究进行了展望,以期为香椿老叶的开发利用提供一定的思路。

生物有机肥对设施香椿产量、品质及土壤养分的影响

通过设施引种栽培试验,研究不同用量生物有机肥作为基肥时对土壤养分及香椿产量、品质的影响。结果表明,与不施生物有机肥相比,施用生物有机肥处理显著增加香椿的分枝数和叶片数,提高香椿嫩芽产量和品质。随着生物有机肥施用量的增加,香椿叶片全氮、全磷、全钾养分含量呈先增加后减少再增加的“N”形趋势,根层0~20cm土层土壤有机质、碱解氮、速效磷、速效钾含量明显提高。综合考虑香椿嫩芽产量、品质、叶片养分积累及土壤肥力变化,推荐生物有机肥施用量为60000~90000kg·hm^(-2)。

基于SSR标记分析浙西南香椿天然居群的遗传多样性

【目的】科学保护和合理利用香椿天然遗传资源,揭示浙西南香椿天然群体的分子遗传背景。【方法】利用16个多态性SSR标记对浙西南5个香椿天然居群的156份香椿材料进行遗传多样性、遗传分化程度和遗传结构分析。【结果】1)156份香椿材料在16个SSR位点共检测出110个等位基因,每个位点的平均等位基因数(Na)和平均有效等位基因数(Ne)分别为6.88和2.48,不同等位基因在香椿个体中呈不均匀分布;16个SSR位点的多态性信息含量(PIC)介于0.04~0.79,平均为0.45;2)16个SSR位点的Shannon’s信息指数(I)和Nei’s基因多样性指数(H)平均值分别为0.99和0.49;居群内近交系数(F_(is))和总近交系数(F_(it))平均值分别为-0.02和0.03,遗传分化系数(F_(st))平均值为0.05,仅5%的遗传变异来自不同居群间;基因流(N_(m))平均值为4.82;分子方差(AMOVA)分析显示遗传分化的根本原因为总的个体内变异(95%),极少为居群间变异(5%),没有来自居群内的个体变异;3)遗传差异和UPGMA聚类分析表明5个居群的Nei’s基因遗传一致度为0.93。浙江开化和浙江常山居群的遗传差异最小,Nei’s基因遗传一致度为0.97;浙江龙游居群与其他4个居群的遗传差异较大;4)群体遗传结构分析表明156份香椿材料无最佳K值;根据ΔK值最大分为3个亚群,各居群材料在3个亚群中皆有一定数量分布。【结论】浙西南天然居群香椿资源在个体水平上的遗传多样性较为丰富;居群内近交程度很低,主要繁殖方式为杂交;香椿居群间的遗传分化程度很小,居群间存在频繁的基因交流,遗传变异主要发生在个体内;浙西南香椿不同居群间的遗传差异总体上较小;居群间未形成相对独立的遗传结构,绝大部分的香椿种质遗传背景比较单一,少量种质拥有复杂的遗传背景。

10年生香椿生长与形质性状的种源变异及选择

【目的】通过分析10年生香椿Toona sinensis生长与形质性状的种源变异,揭示不同性状在地理种源上的变异规律,并进行种源区划及优良种源选择。【方法】以浙江开化县林场的10年生香椿种源林为试材,测定其树高、胸径、枝下高、冠幅、材积、树干圆满度和树冠圆满度,获取各种源生态环境因子,进行方差、主成分、相关性和聚类等分析,计算广义遗传力、综合选择指数和遗传增益,阐明生长和形质性状地理变异模式及与产地生态因子关系。【结果】树高、胸径、树干圆满度和材积存在极显著(P<0.01)种源差异,种源胸径、树高、冠幅、枝下高和材积呈典型的纬向变异模式。种源胸径与冠幅、枝下高和材积之间均呈极显著(P<0.01)正相关。胸径与产地年平均气温、最冷月平均日最低气温和最干燥季节日平均气温等显著(P<0.05)正相关。根据种源聚类结果,可将香椿划分为南部和北部2个种源区,并筛选出3个优良种源,分别为元谋、恩施和太和。【结论】10年生香椿种源间差异显著,并且树高、胸径、材积等性状存在从南向北逐渐减小的变异规律,对其进行选择时胸径是主要的遗传改良性状,可为材用型香椿良种选择提供可靠依据。

不同种源香椿无性系苗期表型性状变异及其与地理-气候因子的相关性

以香椿〔Toona sinensis(A.Juss.)Roem.〕12个种源142个无性系为研究对象,研究了不同种源香椿无性系苗期的表型性状变异,探究其表型性状间及表型性状与地理-气候因子间的相关性。结果表明:香椿无性系苗期表型性状在种源间和种源内差异极显著(P<0.01),且种源内的变异大于种源间的变异。12个种源香椿无性系苗期表型性状的平均变异系数变幅为18.17%~26.34%,变异系数较大。聚类分析结果表明:在欧氏距离17处,供试的12个香椿种源被分成3类,3个类群之间具有明显的表型分化,但聚类结果与地理分布无明显相关性。相关性分析结果表明:香椿无性系苗期的树皮颜色与苗高、地径分别呈显著(P<0.05)和极显著负相关,与叶痕长和叶痕宽的相关性不显著;苗高与经度呈显著负相关,与海拔呈显著正相关;地径与平均气温日较差呈显著负相关,与等温性呈极显著负相关。综上所述,不同种源香椿无性系苗期表型性状变异丰富,表型性状变异主要来源于种源内,具有优良种质资源的选择潜力,且在所收集的种源范围内,苗高的地理变异模式以经度和海拔变异为主。