Main work

In full view of the life cycle, Prof. Li Wanfeng's team found that LaDAL1 transcription was related with life-cycle progression, and its transcript level increased sharply from age 3 to 5 years, which might be the molecular characteristic of the vegetative phase change, and then stayed at a high level. During sexual reproduction, LaDAL1 transcript level decreased sequentially during meiosis and embryogenesis, suggesting that meiosis rapidly lowers the age signal, and after fertilization, the age signal was reset to "0" with the embryogenesis. When a seed germinates, the next generation restarts, and the age is re-counted. Altogether, these results not only provide important and novel insights into the life-cycle progression and trans-generation in perennial woody plants, but also advance our understanding of age recording.

Dormancy release and reactivation of temperate-zone trees involve the temperature-modulated expression of cell-cycle genes. However, information on the detailed regulatory mechanism is limited. Prof. Li Wanfeng's team compared the transcriptomes of the stems of active and dormant larch trees, emphasizing the expression patterns of cell-cycle genes and transcription factors and assessed their relationships and responses to temperatures. They found that twelve cell-cycle genes and 31 transcription factors were strongly expressed in the active stage. Promoter analysis suggested that these 12 genes might be regulated by transcription factors from 10 families. Altogether, 73 cases of regulation between 16 transcription factors and 12 cell-cycle genes were predicted, while the regulatory interactions between LaMYB20 and LaCYCB1; 1, and LaRAV1 and LaCDKB1; 3 were confirmed by yeast one-hybrid and dual-luciferase assays. Last, we found that LaRAV1 and LaCDKB1; 3 had almost the same expression patterns during dormancy release and reactivation induced naturally or artificially by temperature, indicating that the LaRAV1-LaCDKB1; 3 module functions in the temperature-modulated dormancy release and reactivation of larch trees. These results provide new insights into the link between temperature and cell-cycle gene expression, helping to understand the temperature control of tree growth and development in the context of climate change.

Grafting, cutting, and pruning are important horticultural techniques widely used in the establishment of clonal forestry. After the application of these techniques, some properties of the plants change, however, the underlying molecular mechanisms are still unclear. In our previous study, 27 age-related transcripts were found to be expressed differentially between the juvenile vegetative (1 and 2-year-old) and adult reproductive (25- and 50-year-old) phases of Larix kaempferi. We re-analyzed the 27 age-related transcripts, cloned their full-length cDNA sequences, and measured their responses to grafting, cutting, and pruning. After sequence analysis and cloning, 20 transcription factors were obtained and annotated, most of which were associated with reproductive development, and six (LaAGL2-1/LaDAL1, LaAGL2-2, LaAGL2-3, LaSOC1-1, LaAGL11, and LaAP2-2) showed regular expression patterns with L. kaempferi aging. Based on the expression patterns of these transcription factors in L. kaempferi trees subjected to grafting, cutting, and pruning, we concluded that

• cutting and pruning rejuvenate the plants and change their expression, and the effects of cutting on gene expression are detectable within 14 years, although the cutting seedlings are still maturing during these years;

• within three months after grafting, the rootstock is more sensitive to grafting than the scion and readily becomes mature with the effect of the scion, while the scion is not readily rejuvenated by the effect of the rootstock; and

• LaAGL2-2 and LaAGL2-3 are more sensitive to grafting, while LaAP2-2 is impervious to it. These findings not only provide potential molecular markers to assess the state of plants but also aid in studies of the molecular mechanisms of rejuvenation.

Key research projects

• The molecular mechanism regulating somatic embryogenesis by larch miR171-LaSCL6 (2023.01-2026.12)

• Regulation of CDKB and CYCB expression and their functions during larch cambium activity (2018.01-2021.12)

• Effect of age on response of larch to auxin (2013.01-2015.12)

• Regulatory mechanism of miRNA underlying larch phase transition in a whole in a whole rotation period (2014.04-2016.04)

Published articles & books

• Li Xiang-Yi, Ye Zha-Long, Cheng Dong-Xia, Zang Qiao-Lu, Qi Li-Wang, Li Wan-Feng. 2023 LaDAL1 coordinates age and environmental signals in the life cycle of Larix kaempferi. International Journal of Molecular Sciences 24: 426.

• Tao Gui-Yun, Xie Yun-Hui, Li Wan-Feng, Li Kui-Peng, Sun Chao, Wang Hong-Ming, Sun Xiao-Mei. 2023 LkARF7 and LkARF19 overexpression promote adventitious root formation in a heterologous poplar model by positively regulating LkBBM1. Communications Biology 6.

• Li Xiang-Yi, Cheng Dong-Xia, Qi Li-Wang, Zhan Jin-Jei, Li Wan-Feng. 2023 Regulation of age-dependent expression patterns of five transcription factors in Larix kaempferi. 

• Li Wan-Feng, Qi Li-Wang, Yang Wen-Hua. 2022 Age-related miRNA-mediated regulatory networks orchestrating chronological development of meristems in Larix kaempferi. Journal of Plant Growth Regulation 41: 2305-2318.

• Ye Zha-Long, Zang Qiao-Lu, Cheng Dong-Xia, Li Xiang-Yi, Qi Li-Wang, Li Wan-Feng 2022 Over-expression of larch dal1 accelerates life-cycle progression in Arabidopsis. Forests 13: 953.

• Zang Qiao-Lu, Zhang Yao, Han Su-Ying, Li Wan-Feng, Qi Li-Wang. 2021 Transcriptional and post-transcriptional regulation of the miR171-LaSCL6 module during somatic embryogenesis in Larix kaempferi. Trees 35: 145-154.

• Li Wan-Feng, Kang Yanhui, Zhang Yao, Zang Qiao-Lu, Qi Li-Wang. 2021 Concerted control of the LaRAV1-LaCDKB1; 3 module by temperature during dormancy release and reactivation of larch. Tree Physiology 41: 1918-1937.

• Fan Yan-Ru, Li Wan-Feng, Li Zhe-Xin, Dang Shao-Fei, Han Su-Ying, Zhang Li-Feng, Qi Li-Wang. 2021 Examination of the transcriptional response to LamiR166a overexpression in Larix kaempferi (Lamb.) Carr. Biology 10: 576.

• Ren Qiu-Rong, Sretenovic Simon, Liu Shi-Shi, Tang Xu, Huang Lan, He Yao, Liu Li, Guo Ya-Chong, Zhong Zhao-Hui, Liu Guan-Qing, Cheng Yan-Hao, Zheng Xue-Lian, Pan Chang-Tian, Yin De-Suo, Zhang Ying-Xiao, Li Wan-Feng, Qi Li-Wang, Li Cheng-Hao, Qi Yi-Ping, Zhang Yong. 2021 PAM-less plant genome editing using a CRISPR-SpRY toolbox. Nature Plants 7: 25-33.

• Kang Yan-Hui, Li Wan-Feng, Zhang Li-Feng, Qi Li-Wang. 2021 Over-expression of the cell-cycle gene LaCDKB1; 2 promotes cell proliferation and the formation of normal cotyledonary embryos during Larix kaempferi somatic embryogenesis. Genes 12: 1435.

• Zhang Yao, Zang Qiao-Lu, Qi Li-Wang, Han Su-Ying, Li Wan-Feng. 2020 Effects of cutting, pruning, and grafting on the expression of age-related genes in Larix kaempferi. Forests 11: 218.

• Li Hui, Huang Xiong, Li Wan-Feng, Lu Yan, Dai Xin-Ren, Zhou Zai-Zhi, Li Quan-Zi. 2020 MicroRNA comparison between poplar and larch provides insight into the different mechanism of wood formation. Plant Cell Reports 39: 1199-1217.

• Zang Qiao-Lu, Li Xiang-Yi, Qi Li-Wang, Li Wan-Feng. 2020 Identification and characterization of LaSCL6 alleles in Larix kaempferi (Lamb.) Carr. Based on analysis of simple sequence repeats and allelic expression. Forests 11: 1-10.

• Zang Qiao-Lu, Li Wan-Feng, Qi Li-Wang. 2019 Regulation of LaSCL6 expression by genomic structure, alternative splicing, and microRNA in Larix kaempferi. Tree Genetics & Genomes 15.

• Xiang Wei-Bo, Li Wan-Feng, Zhang Shou-Gong, Qi Li-Wang. 2019 Transcriptome-wide analysis to dissect the transcription factors orchestrating the phase change from vegetative to reproductive development in Larix kaempferi. Tree Genetics & Genomes 15.

• Li Zhe-Xin, Fan Yan-Ru, Dang Shao-Fei, Li Wan-Feng, Qi Li-Wang, Han Su-ying. 2018 LaIR166a-mediated auxin biosynthesis and signalling affect somatic embryogenesis in Larix leptolepis. Molecular Genetics and Genomics 293: 1355-1363.

• Li Wan-Feng, Yang Wen-Hua, Zhang Shou-Gong, Han Su-Ying, Qi Li-Wang. 2017 Transcriptome analysis provides insights into wood formation during larch tree aging. Tree Genetics & Genomes 13: 19.

• Li Zhe-Xin, Zhang Li-Feng, Li Wan-Feng, Qi Li-Wang, Han Su-Ying. 2017 MiR166a affects the germination of somatic embryos in Larix leptolepis by modulating IAA biosynthesis and signaling genes. Journal of Plant Growth Regulation 36: 889-896.

• Li Zhe-Xin, Li Shui-Gen, Zhang Li-feng, Han Su-ying, Li Wan-Feng, Xu Hai-yan, Yang Wen-hua, Liu Ya-li, Fan Yan-ru, Qi Li-Wang. 2016 Over-expression of MIR166a inhibits cotyledon formation in somatic embryos and promotes lateral root development in seedlings of Larix leptolepis. Plant Cell, Tissue and Organ Culture (PCTOC) 127: 461-473.

• Liu Ya-Li, Han Su-Ying, Ding Xiang-Ming, Li Xin-Min, Zhang Li-Feng, Li Wan-Feng, Xu Hai-Yan, Li Zhe-Xin, Qi Li-Wang. 2016 Transcriptome analysis of mRNA and miRNA in somatic embryos of Larix leptolepis subjected to hydrogen treatment. International Journal of Molecular Sciences 17: 1951.

• Wu Bi-fei, Li Wan-feng, Xu Hai-yan, Qi Li-wang, Han Su-ying. 2015 Role of cin-miR2118 in drought stress responses in Caragana intermedia and tobacco. Gene.

• Li Wan-Feng, Zhang Shou-Gong, Han Su-Ying, Wu Tao, Zhang Jun-Hong, Qi Li-Wang. 2014 The post-transcriptional regulation of LaSCL6 by miR171 during maintenance of embryogenic potential in Larix kaempferi (Lamb.) Carr. Tree Genetics & Genomes 10: 223-229.

• Zhang Jun-Hong, Zhang Shou-Gong, Li Shui-Gen, Han Su-Ying, Li Wan-Feng, Li Xin-Min, Qi Li-Wang. 2014 Regulation of synchronism by abscisic-acid-responsive small noncoding rnas during somatic embryogenesis in larch (Larix leptolepis). Plant Cell, Tissue and Organ Culture (PCTOC) 116: 361-370.

• Zhang Li-feng, Li Wan-feng, Xu Hai-yan, Qi Li-wang, Han Su-ying. 2014 Cloning and characterization of four differentially expressed cDNAs encoding NFYA homologs involved in responses to ABA during somatic embryogenesis in Japanese larch (Larix leptolepis). Plant Cell, Tissue and Organ Culture (PCTOC) 117: 293-304.

• Li Wan-Feng, Han Su-Ying, Qi Li-Wang, Zhang Shou-Gong. 2014 Transcriptome resources and genome-wide marker development for Japanese larch (Larix kaempferi). Frontiers of Agricultural Science and Engineering 1: 77.

• Zhang Li-feng, Li Wan-feng, Han Su-ying, Yang Wen-hua, Qi Li-wang. 2013 CDNA cloning, genomic organization and expression analysis during somatic embryogenesis of the translationally controlled tumor protein (TCTP) gene from Japanese larch (Larix leptolepis). Gene 529: 150-158.

• Li Shui-gen, Li Wan-feng, Han Su-ying, Yang Wen-hua, Qi Li-wang. 2013 Stage-specific regulation of four HD-ZIP Ⅲ transcription factors during polar pattern formation in Larix leptolepis somatic embryos. Gene 522: 177-183.

• Li Wan-Feng, Ding Qi, Cui Ke-Ming, He Xin-Qiang. 2013 Cambium reactivation independent of bud unfolding involves de novo IAA biosynthesis in cambium regions in Populus tomentosa carr. Acta Physiologiae Plantarum 35: 1827-1836.

• Li Wan-Feng, Zhang Shou-Gong, Han Su-Ying, Wu Tao, Zhang Jun-Hong, Qi Li-Wang. 2013 Regulation of LaMYB33 by miR159 during maintenance of embryogenic potential and somatic embryo maturation in Larix kaempferi (Lamb.) Carr. Plant Cell, Tissue and Organ Culture (PCTOC) 113: 131-136.

• Zhu Jian-Feng, Zhang Li-Feng, Li Wan-Feng, Han Su-Ying, Yang Wen-Hua, Qi Li-Wang. 2013 Reference gene selection for quantitative real-time PCR normalization in Caragana intermedia under different abiotic stress conditions. Plos One 8:   e53196.

• Zhu Jian-Feng, Li Wan-Feng, Yang Wen-Hua, Qi Li-Wang, Han Su-Ying. 2013 Identification of microRNAs in Caragana intermedia by high-throughput sequencing and expression analysis of 12 microRNAs and their targets under salt stress. Plant Cell Reports 32: 1339-1349.

• Zhang Shou-Gong, Han Su-Ying, Li Wan-Feng, Zhou Jian, Li Xin-Min, Qi Li-Wang. 2012 Mirna regulation in fast- and slow-growing hybrid Larix trees. Trees 26: 1597-1604.

• Zhang Jun-Hong, Zhang Shou-Gong, Han Su-Ying, Wu Tao, Li Xin-Min, Li Wan-Feng, Qi Li-Wang. 2012 Genome-wide identification of microRNAs in larch and stage-specific modulation of 11 conserved microRNAs and their targets during somatic embryogenesis. Planta 236: 647-657.

• Li Wan-Feng, Cui Ke-Ming, He Xin-Qiang. 2009 Regulation of cell cycle regulators by environmental signals during growth-dormancy cycle of trees. Plant Signaling & Behavior 4: 959-961.

• Li Wan-Feng, Ding Qi, Chen Jia-Jia, Cui Ke-Ming, He Xin-Qiang. 2009 Induction of PtoCDKB and PtoCYCB transcription by temperature during cambium reactivation in Populus tomentosa carr. Journal of Experimental Botany 60: 2621-2630.

• Zhang Shou-Gong, Zhou Jian, Han Su-Ying, Yang Wen-Hua, Li Wan-Feng, Wei Hua-Li, Li Xin-Min, Qi Li-Wang. 2010 Four abiotic stress-induced mirna families differentially regulated in the embryogenic and non-embryogenic callus tissues of Larix leptolepis. Biochemical and Biophysical Research Communications 398: 355-360.

• Hou Hong-Wei, Zhou Yu-Ting, Mwange Kalima-N'Koma, Li Wan-Feng, He Xin-Qiang, Cui Ke-Ming. 2006 ABP1 expression regulated by IAA and ABA is associated with the cambium periodicity in Eucommia ulmoides oliv. Journal of Experimental Botany 57: 3857-3867.