Subjects: Biology >> Botany >> Applied botany submitted time 2023-07-26 Cooperative journals: 《广西植物》
Abstract： In order to select superior clones with strong resin-producing capacity and high resin-quality from the existing fast-growing timber clones in seed orchard of Pinus elliottii, and to make high use of crop germplasm resource,36 clones in the first generation of slash pine seed orchard were used as materials to determine their resin yield, resin mass flow rate and DBH growth, and to analyze their resin composition by GC-MS. Based on the above indicators, correlation analysis and cluster analysis was used to comprehensively evaluate the production and quality of resin among 36 clones. The results were as follows: (1) There were a total of 21 pine resin components, including 8 monoterpenes and 13 diterpenes. (2) Correlation analysis showed that the resin mass flow rate (RMR) had significant and positive correlation with the total content of monoterpene, weakly negatively correlates to abietic-type resin acid, and not significantly correlated to pimaric-type resin acid. (3) Based on the cluster analysis results integrating four types of indicators as the total monoterpene content, resin mass flow rate, abietic-type resin acid and pimaric-type resin acid, 36 clones could be divided into three categories and the difference between each type is significant. The performance of Class 1 was much better than that of the other two categories. (4) There were 17 high-resin yield pine clones (ERM≥15.15) among 36 clones, and on the basis of this, four clones (6-44, 4-11-1, 1-38 and 3-64) display higher monoterpenes content, while four clones (4-11-1, 3-64, 2-0420, 3-468) showed higher contents of pimaric-type resin acid. And the content of abietic-type resin acid of clone 2-173 was the highest. In summary, a total of 21 pine resin components of P. elliottii were identified, and 36 clones were evaluated based on four indicators: the total monoterpene content, resin mass flow rate, abietic-type resin acid and pimaric-type resin acid. We not only analyzed qualitatively the resin composition but also evaluated quantitatively the resin-producing capacity of 36 clones in slash pine seed orchard. Our findings provide the scientific references for the targeted breeding of pine resin components and lay a foundation for subsequent heredity breeding and gene improvement of P. elliottii.
Subjects: Biology >> Botany >> Applied botany submitted time 2021-07-20 Cooperative journals: 《广西植物》
Abstract： AGO1 plays an important role in the differentiation of leaf primordia, originating from the the peripheral region of Shoot Apical Meristem(SAM). To study the morphogenesis mechanism of leaf primordium differentiation, we cloned 2 001bp upstream region of LtAGO1 CDS as the promoter by RT-PCR and RACE technology on the basis of previous cloned LtAGO1 gene sequence, and predicted its function. Real-time PCR was used to investigate expression pattern in Liriodendron L. We obtained the transgenic Arabidopsis thaliana of ProAGO1::GUS by resistance screening and DNA dentification, and then monitored phenotype and GUS histochemical staining. The results were as follows: (1) The LtAGO1 gene included an open reading frame for 3300 bp , encoded 1 100 amino acid , the molecular weight was 122.14 kD and theoretical isoelectric point was 9.36. (2)Amino acid sequence analysis showed that it consisted of Gly-rich-AGO1 and Piwi conserved domains of AGO family. Phylogenetic trees revealed that LtAGO1 was closed to Cinnamomum micranthum （RWR84608.1） in evolutional relationship. (3)The specific tissues expression analysis demonstrated that the expression order was that stamen>floral bud>petal>calyx>leaf>pistil>leaf bud>stem among tissues, and the expression order was that leaf bud sprouting stage >young leaf stage>senescence stage >mature stage among stages. It was highly expressed in the leaf margin of Liriodendron L, and LtAGO1 gene expression in leaf tooth sinus was higher than in leaf tooth tip of Liriodendron tulipifera. (4)The transgenic strains leaf polarity of the middle and basal apical axis was absent with serrated leaf margin and double petal flower. It was found that GUS staining was stably detected at the tip of leaf bud of transgenic seedlings , the higher GUS activity was observed at newly differentiated petioles. LtAGO1 promoter drove GUS gene to accumulate specifically in the vascular bundle of Arabidopsis thaliana leaf , flower, pod and stem, and GUS activity intensity order was that leaf tip bud> flower>vascular bundle among tissues, which was accordance with the Real-time PCR results in Liriodendron tulipifera. Therefore, the results also showed that LtAGO1 gene was predominantly expressed in apical meristem and regulated by various pathways during the development of leaf and flower. It will provide a foundation for further functional research of AGO1 protein and regulation mechanism of leaf shape development.
Subjects: Biology >> Botany >> Applied botany submitted time 2020-08-03 Cooperative journals: 《广西植物》
Abstract：法尼基焦磷酸合酶（farnesyl diphosphate synthase, FPPS）是植物萜类物质合成前体法尼基焦磷酸（farnesyl pyrophosphate，FPP）的关键合成酶。为探究北美鹅掌楸（Liriodendrontulipifera）萜类合成途径的分子机制，该研究利用北美鹅掌楸转录组数据，通过设计特异性引物，采用 RACE 技术克隆得到 LtuFPPS1 基因的全长序列并进行生物信息学分析。结果表明：（1）LtuFPPS1 基因全长 1 460 bp，开放阅读框（ORF）长 1 056 bp，编码 351 个氨基酸，蛋白质分子量为 40 589.45 D，等电点为 5.19，不稳定系数为 43.50，归类为不稳定蛋白；LtuFPPS1 基因所编码的蛋白不含信号肽，定位于线粒体，是一种不稳定亲水性蛋白，具有类异戊二烯类化合物合酶的特征结构域，α-螺旋为其氨基酸序列的主要二级结构。（2）进化树和同源序列分析表明，北美鹅掌楸 LtuFPPS1 蛋白与乐昌含笑（Michelia chapensis）的 FPPS蛋白的亲缘关系更近。（3）组织表达分析结果发现，LtuFPPS1 基因在北美鹅掌楸的雌蕊中的表达量最高，其高低顺序为雌蕊 > 花芽 > 茎 > 雄蕊 > 萼片 > 叶片 > 花瓣 > 根，据此推测萜类代谢物在花器官中的合成相对较多。综上所述，LtuFPPS1 基因为萜类合成酶基因，可以为从分子生物学层面探究北美鹅掌楸萜类物质的合成提供一定的理论帮助。
Subjects: Biology >> Botany >> Applied botany submitted time 2020-01-12 Cooperative journals: 《广西植物》
Abstract：为使速生湿地松良种快速大规模繁殖，对其胚性愈伤组织进行诱导和增殖优化研究。该研究以1代湿地松种子园中10个速生湿地松优良无性系（基因型）的未成熟合子胚为外植体，系统研究了基因型、合子胚发育阶段、基本培养基、植物生长调节剂种类和浓度等不同因子对胚性愈伤组织诱导效率的影响，并探讨了胚性愈伤组织的增殖条件。结果表明：基因型、合子胚发育阶段、基本培养基及植物生长调节剂种类和浓度等均对胚性愈伤组织诱导有不同程度的影响。10个基因型均诱导出了胚性愈伤组织，其中以2号基因型诱导率最高，达到25.37%；合子胚发育阶段中，以处于多胚阶段的球果诱导率最高；5种基本培养基中，以在DCR基本培养基上诱导效果最佳；植物生长调节剂浓度及组合中以2,4-D 2.0 mg·L-1 + KT 2.5 mg·L-1诱导率最高，达到27.78%。湿地松胚性愈伤组织增殖的最佳培养基为在DCR基本培养基上添加0.5 mg·L-1 2,4-D、1.0 mg·L-1 KT、500 mg·L-1 CH和300 mg·L-1 L-谷氨酰胺。该研究为湿地松良种进一步开展成熟胚诱导及植株再生奠定了基础。
Subjects: Biology >> Botany >> Applied botany submitted time 2019-08-27 Cooperative journals: 《广西植物》
Abstract：交替氧化酶（AOX）为植物呼吸作用电子传递链末端氧化酶，广泛存在于高等植物中，研究表明AOX在植物生长发育及适应性方面有着重要作用。鹅掌楸（Liriodendron chinense）为重要的珍贵用材及园林观赏树种，开展抗逆基因的研究，对于提高鹅掌楸适应性有重要意义。该文以鹅掌楸为研究对象，通过采用RT-PCR与RACE相结合的方法克隆获得3个AOX基因，其ORF长分别为858 bp、1 032 bp和1 044 bp，相应编码氨基酸数为285 aa、343 aa和347 aa，分别命名为LcAOX1a、LcAOX1b和LcAOX2。蛋白同源性分析发现鹅掌楸AOX家族蛋白序列高度保守，尤其在C端保守性极高，且均含有“EXXH”、“EEE-Y” 铁离子结合保守结构域。亚细胞定位分析结果显示LcAOX1a蛋白定位于线粒体及叶绿体之外的其他位置，LcAOX1b蛋白在叶绿体和线粒体中均有定位，LcAOX2蛋白定位于线粒体基质。采用RT-qPCR方法研究AOX基因在鹅掌楸茎、叶片、叶芽、花芽、花萼、花瓣、雄蕊、雌蕊8个不同组织中的表达模式，分析发现鹅掌楸AOX基因在花器官中表达量明显高于营养器官，LcAOX1a与LcAOX1b基因在雄蕊中表达量最高，特别是LcAOX1a基因在雄蕊中特异性表达，其表达量远远高于其他组织；而LcAOX2基因在花瓣中表达量最高。该研究克隆3个AOX基因并进行相关分析，为进一步研究其生物学功能奠定基础。
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