Abstract The OVATE family protein (OFP) is a plant-specific regulatory protein that plays a significant role in regulating plant morphogenesis and secondary cell wall formation. However, little information about the OFP gene family in Chinese fir (Cunninghamia lanceolata) is available. In this study, ClOFP genes were identi-fied based on the full-length transcriptome. Bioinformatics characteristics and evolutionary relationships were analyzed using online service. Besides, relative expression patterns of all members in different tissues and or-gans were determined by real time fluorescence quantitative PCR (qRT-PCR). Furthermore, the subcellular lo-calizations of all proteins were determined by transient transfection assays. The results showed that 6 ClOFP genes were identified from RNA-seq data, with a typical OVATE domain and contained no introns. The phylo-genetic analysis revealed that ClOFP were distributed in 5 subgroups. In addition, relative expression patterns of different tissues and organs varied differently. ClOFP1 was strongly expressed in young leaves and the rela-tive expressions in stems were approximately decreased with an increase of lignification. ClOFP2 dominantly expressed in female cone. ClOFP3 strongly expressed in root. The relative expression of ClOFP2/3 in low lig-nified stem (S1) was higher than that in others. ClOFP4 dominantly expressed in female cone, male cone and leaves. ClOFP5 was strongly expressed in male cone and the relative expressions of ClOFP5 in stems were ap-proximately increased with an increase of lignification. ClOFP6 was strongly expressed in stems and female cone. Moreover, the relative expressions of ClOFPs in leaves were decreased with an increase in maturity. The results of subcellular localization showed the localization of ClOFP1 in the cytoplasm, ClOFP2/3 in the plas-ma membrane and nuclear localization of ClOFP4/5/6. This suggested that ClOFPs might involve in different regulatory pathways in Chinese fir. Hence, ClOFPs might involve in regulating the development of cone, leaf, stem and other organs, as well as lignin synthesis. The research results provides a crucial theoretical basis for further research on organogenesis and wood formation of Chinese fir, At the same time, it also provides poten-tial genetic resources for molecular breeding of C. lanceolata.
YING Wei-Yang,HU Xian-Ge,HUANG Hua-Hong, et al. Expression Pattern and Subcellular Localization of the OFP Gene Family in Chinese Fir (Cunninghamia lanceolata)[J]. 农业生物技术学报, 2023, 31(8): 1610-1622.
[1] 程健弘, 魏明科, 林二培, 等. 2017. 杉木 HD-Zip Ⅲ转录因子的克隆及表达分析[J]. 农业生物技术学报, 25(11):1820-1830. (Cheng J H, Wei M K, Lin E P, et al. 2017. Cloning and expression analysis of HD-Zip Ⅲ transcriptional factors in Cunninghamia lanceolata[J]. Journal of Agricultural Biotechnology, 25(11): 1820-1830.) [2] 郭昱, 李娜, 李珊, 等. 2020. 谷子 OFP 基因家族的鉴定及表达分析[J]. 山西农业科学, 48(4): 467-476. (Guo Y, Li N, Li S, et al. 2020. Identification and ex-pression analysis of OFP gene family in Setavia italica [J]. Journal of Shanxi Agricultural Sciences, 48(4):467-476.) [3] 李树斌, 周丽丽, 闫新阳, 等. 2020. 杉木 CL14-3-3-e 基因克隆及其在干旱胁迫下表达分析[J]. 分子植物育种, 18(16): 5306-5314. (Li S B, Zhou L L, Yan X Y, et al. 2020. Gene cloning and its expression analysis under drought-stress of CL14-3-3-e of Chinese fir[J]. Molecu-lar Plant Breeding, 18(16): 5306-5314.) [4] 李振兴, 单雪梅, 王国栋, 等. 2020. 杉木属植物的化学成分及药理作用研究进展[J]. 特产研究, 42(6): 79-84. (Li Z X, Shan X M, Wang G D, et al. 2020. Research prog-ress on chemical constituents and pharmacological ef-fects of Cunninghamia[J]. Special Wild Economic Ani-mal and Plant Research, 42(6): 79-84.) [5] 齐薇娜, 罗智, 李洪太, 等. 2021. 枣 OVATE 基因家族的鉴定与生物信息学分析[J]. 分子植物育种, 19(7): 2207-2216. (Qi W N, Luo Z, Li H T, et al. 2021. Identification and bioinformatics analysis of OVATE gene family in Chinese jujube[J]. Molecular Plant Breeding, 19(7):2207-2216.) [6] 苏烁烁, 李明, 吴鹏飞, 等. 2017. 杉木磷转运蛋白基因ClPht1; 1 的克隆及表达分析[J]. 林业科学, 53(5): 33-42. (Su S S, Li M, Wu P F, et al. 2017. Cloning and ex-pression analysis of phosphate transporter gene ClPht1;1 in Cunninghamia lanceolata[J]. Scientia Silvae Sini-cae, 53(5): 33-42.) [7] 王静雅, 刘文娟, 吕祎馨, 等. 2020. 油桐 OFP 基因家族的全基因组鉴定及低温响应分析[J]. 植物生理学报, 56(5):949-960. (Wang J Y, Liu W J, Lv Y X, et al. 2020. Ge-nome-wide identification and expression analysis under-coldstress of OFP gene family in Vernicia fordii[J]. Plant Physiology Journal, 56(5): 949-960.) [8] 王钰, 童再康, 黄华宏, 等. 2010. 光皮桦总 RNA 的提取及 Actin 基因的克隆[J]. 浙江林业科技, 30(4): 32-36. (Wang Y, Tong Z K, Huang H H, et al. 2010. Extracting of total RNA from Betula luminifera and the cloning of Actin gene[J]. Journal of Zhejiang Forestry Science and Technology, 30(4): 32-36.) [9] 伍汉斌, 段爱国, 张建国. 2019. 杉木地理种源不同林龄生长变异及选择[J]. 林业科学, 55(10): 181-192. (Wu H B, Duan A G, Zhang J G, 2019. Growth variation and selection effect of Cunninghamia lanceolata provenances at different stand ages[J]. Scientia Silvae Sinicae, 55(10):181-192.) [10] 谢一青, 李志真, 黄儒株, 等. 2006. 光皮桦基因组 DNA 提取方法比较[J]. 浙江林学院学报, 23(6): 664-668. (Xie Y Q, Li Z Z, Huang R Z, et al. 2006. Comparison of meth-ods of extracting genomic DNA from Betula luminifera [J]. Journal of Zhejiang Forestry College, 23(6): 664-668.) [11] 许瑞瑞, 李睿, 王小非, 等. 2018. 苹果 OFP 基因家族的全基因组鉴定与非生物逆境表达分析[J]. 中国农业科学,51(10): 1948-1959. (Xu R R, Li R, Wang X F, et al. 2018. Identification and expression analysis under abiot-ic stresses of OFP gene family in apple[J]. Scientia Agri-cultura Sinica, 51(10): 1948-1959.) [12] 张彦琴, 常建忠, 董春林, 等. 2020. 玉米 OFP 基因家族全基因组鉴定及表达分析[J]. 分子植物育种, 18(20): 6595-6604. (Zhang Y Q, Chang J Z, Dong C L, et al. 2020. Genome-wide identification and expression analysis of OFP gene family in Zea mays[J]. Molecular Plant Breed-ing, 18(20): 6595-6604.) [13] 赵林峰, 高建亮. 2021. 杉木无性系不同林龄生长变异与选择效应[J]. 西北农林科技大学学报, 50(1): 1-9. (Zhao L F, Gao J L.2021. Growth variation and selection ef-fect of Cunninghamia lanceolata clones at different stand ages[J]. Journal of Northwest A&F University, 50(1): 1-9.) [14] Chen H Y, Yu H, Jiang W Z, et al. 2021. Overexpression of OVATE family protein 22 confers multiple morphologi-cal changes and represses gibberellin and brassino-steroid signalings in transgenic rice[J]. Plant Science, 304: 110734. [15] Ku H M, Doganlar S, Chen K Y, et al. 1999. The genetic basis of pear-shaped tomato fruit[J]. Theoretical and Applied Genetics, 99(5): 844-850. [16] Li E Y, Wang S C, Liu Y Y, et al. 2011. Ovate family protein4 (OFP4) interaction with KNAT7 regulates secondary cell wall formation in Arabidopsis thaliana[J]. Plant Jour-nal, 67(2): 328-341. [17] Liu D, Sun W, Yuan Y W, et al. 2014. Phylogenetic analyses provide the first insights into the evolution of OVATE family proteins in land plants[J]. Annals of Botany, 113(7): 1219-1233. [18] Liu J P, Van Eck J, Cong B, et al. 2002. A new class of regula-tory genes underlying the cause of pear-shaped tomato fruit[J]. Proceedings of the National Academy of Scienc-es of the USA, 99(20): 13302-13306. [19] Ma Y M, Yang C, He Y, et al. 2017. Rice OVATE family pro-tein 6 regulates plant development and confers resis-tance to drought and cold stresses[J]. Journal of Experi-mental Botany, 68(17): 4885-4898. [20] Nystedt B., Street N., Wetterbom A,et al. 2013. The Norway spruce genome sequence and conifer genome evolution[J]. Nature, 497: 579-584. [21] Pagnussat G C, Yu H J, Sundaresan V.2007. Cell-fate switch of synergid to egg cell in Arabidopsis eostre mutant em-bryo sacs arises from misexpression of the BEL1-like homeodomain gene BLH1[J]. Plant Cell, 19(11): 3578-3592. [22] Schmitz A J, Begcy K, SaratH G, et al. 2015. Rice OVATE family protein 2 (OFP2) alters hormonal homeostasis and vasculature development[J]. Plant Science, 241:177-188. [23] Sun X X, Ma Y M, Yang C, et al. 2020. Rice OVATE family protein 6 regulates leaf angle by modulating secondary cell wall biosynthesis[J]. Plant Molecular Biology, 104(3): 249-261. [24] Tang Y, Zhang W, Yin Y L, et al. 2018. Expression of OVATE family protein 8 affects epicuticular waxes accumulation in Arabidopsis thaliana[J]. Botanical Studies, 59(1):1-9. [25] Wang D D, Hao Z D, Long X F, et al. 2020. The transcrip-tome of Cunninghamia lanceolata male/female cone re-veal the association between MIKC MADS-box genes and reproductive organs development[J]. BMC Plant Bi-ology, 20(1): 1-12. [26] Wang R H, Han T L, Sun J F, et al. 2021. Genome-wide identi-fication and characterization of the OFP gene family in Chinese cabbage (Brassica rapa L. ssp. pekinensis)[J]. PeerJ, 9: e10934. [27] Wang S C, Chang Y, Guo J J, et al. 2007. Arabidopsis OVATE family protein 1 is a transcriptional repressor that sup-presses cell elongation[J]. The Plant Journal, 50(5): 858-872. [28] Wang S C, Chang Y, Guo J J, et al. 2011. Arabidopsis OVATE family proteins, a novel transcriptional repressor family, control multiple aspects of plant growth and develop-ment[J]. PLOS ONE, 6(8): e23896. [29] Yang C, Ma Y M, He Y, et al. 2018. OsOFP19 modulates plant architecture by integrating the cell division pattern and brassinosteroid signaling[J]. The Plant Journal, 93(3): 489-501. [30] Yu H, Jiang W Z, Liu Q, et al. 2015. Expression pattern and subcellular localization of the OVATE protein family in rice[J]. PLOS ONE, 10(3): 1-19.
