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  • Hosted by:Chinese Academy of Sciences
    Sponsored by:Institute of Botany, Chinese Academy of Sciences, Botanical Society of China
    Co-hosted by:Key Laboratory of Soybean Molecular Design Breeding, Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences
    Institute of Biotechnology and Germplasm Resources, Yunnan AgriculturalAcademy
    Fujian Agriculture and Forestry University
    Hunan Provincial Key Laboratory of Phytohormones and Growth Development, Hunan Agricultural University
    State Key Laboratory of Crops Biology, Shandong Agricultural University

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Current Issue
Editor-in-Chief:Lei Wang
ISSN 1674-3466 CN 11-5705/Q
Post Code:2-967
Volume 61 Issue 3
10 May 2026
  
SPOTLIGHT
Precision Engineering of Strigolactone Receptor Aids Rice in Antiviral Defense
Yang Jinhua, Wang bing
Chinese Bulletin of Botany. 2026, 61(3):  357-368.  doi: 10.3724/CBB-2026-0044
Abstract ( 72 )   PDF (1383KB) ( 29 )   Save
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The RNA interference (RNAi) pathway acts as a conserved and broad-spectrum antiviral defense mechanism in plants, whereas viruses deploy effector proteins to counter host defense. Strigolactones (SLs) are a new class of phytohormones that regulate plant architecture, stress resilience, arbuscular mycorrhizal symbiosis, and parasitic diseases, but their antiviral mechanisms remain unclear. Recent research reveals that SL signaling is a pivotal regulatory node in the arms race between viruses and plants. SLs induce the expression of the transcription factor ONAC131, which acts synergistically with the virus-responsive protein MID1 to enhance antiviral RNAi. However, the rice grassy stunt virus (RGSV) deploys the P3 effector to dampen SL perception by disrupting the interaction between the SL receptor D14 and D3, thereby weakening the antiviral defense of rice. Precise editing of D14 remarkably enhances resistance to RGSV without yield penalty. This work reveals a novel mechanism whereby the SL signaling pathway and the RNAi pathway cooperate to counter viral invasion. It also provides a precise target for antiviral breeding in crops and represents a major breakthrough in the field of plant virology. In light of this, this paper summarizes the research progress on the antiviral RNAi pathway and SL signaling pathway, highlighting the regulatory functions of SL and other phytohormone signaling pathways in the offensive and defensive interactions between plants and viruses. Looking ahead, it offers new insights for the targeted breeding of antiviral crops by identifying key genes in hormone signaling hubs.


INVITED REVIEWS

The Regulatory Network and Mechanism of Plant Floral Organ Development Mediated by Auxin Response Factors

Xinyu Liu, Silan Dai
Chinese Bulletin of Botany. 2026, 61(3):  369-385.  doi: 10.11983/CBB25169
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Auxin response factors (ARFs), as core transcription factors in auxin signal transduction, are deeply involved in the entire process of auxin-regulated plant floral organ development. This review summarizes the key roles of ARFs in regulating floral organ differentiation (such as floral primordium initiation and meristem maintenance) and morphogenesis (development of sepals, petals, stamens, carpels, and regulation of organ boundaries) by responding to auxin signals. It also elaborates on the coordinated regulatory mechanism between ARFs and MADS-box genes in the ABCDE model, and reveals the molecular basis of the network formed through post-transcriptional regulation by miRNAs and interactions with hormone signals such as cytokinins, which together maintain the homeostasis of floral development. This provides a theoretical basis for analyzing the molecular mechanism of plant floral development and offers a theoretical foundation for molecular breeding of flowers.
RESEARCH ARTICLE
Construction of Yeast Two-hybrid cDNA Library and Screening of Interacting Proteins of SOC1a in Soybean Shoot Apexes and Axillary Buds
Huan Huang, Jiali Zhang, Xue Yang, Liyu Chen, Lin Yue, Baohui Liu , Hui Yang
Chinese Bulletin of Botany. 2026, 61(3):  386-401.  doi: 10.11983/CBB25062
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INTRODUCTION: The shoot apexes and axillary buds determine crop growth and yield potential, with their developmental states directly shaping shoot architecture. However, there are currently few cDNA libraries constructed for shoot apexes and/or axillary buds in soybean (Glycine max).


RATIONALE: By constructing cDNA libraries for shoot apexes and axillary buds, we can gain an in-depth understanding of the core mechanisms underlying plant architecture in soybean at the molecular level, thereby providing theoretical foundations and genetic resources for the design of high-yielding and well-adapted soybean varieties.


RESULTS: This study constructed a yeast two-hybrid (Y2H) nuclear system cDNA library using shoot apexes and axillary buds from the cultivar "Williams 82" grown under long-day and short-day conditions at different developmental stages. Equal amounts of RNA extracted from these tissues were pooled and subjected to cDNA library construction using the Gateway method, followed by transcript diversity analysis. The resultant library had a capacity of 1.2×10⁷ CFU, with 100% recombination rate and an average length exceeding 1000 bp of the inserted fragments, covering 29,170 genes. This cDNA library meets the library construction standards and is suitable for subsequent Y2H screening. Using the key floral transition and shoot architecture regulator SOC1a as a bait, we first tested the toxicity and self-activation of the recombinant pGBKT7-SOC1a and then performed library screening. A total of 50 positive clones were obtained, and after DNA sequencing, BLAST alignment, and functional annotation, 14 candidate interacted proteins were identified. Among them, five candidate proteins were cloned into pGADT7 vector and subjected to pairwise retransformation assays with pGBKT7-SOC1a, confirming physical interactions between two of these proteins and SOC1a. Furthermore, the interaction between SOC1a and one of the candidate proteins, SEP2, was demonstrated through co-immunoprecipitation and luciferase complementation imaging assays.


CONCLUSION: This study establishes a high-quality Y2H cDNA library for soybean meristematic tissues and identifies novel SOC1a-interacting proteins, providing critical molecular insights into SOC1a-mediated regulation of soybean shoot architecture development.




Screening of SOC1a-interacting proteins using a yeast cDNA library constructed from soybean shoot apexes and axillary buds (A), with interactions confirmed by yeast retransformation (B), co-immunoprecipitation (C), and luciferase complementation imaging (D) assays.



Molecular Mechanisms of Glycosyltransferases CsUGT73B4 and CsUGT85K5 in Tea Plants in Response to Cold Stress
Gai Xinyue, Wang Xiaodong, Fan Yangen, Li Bin, Fu Xiaodong, Sun Ping, Huang Xiaoqin
Chinese Bulletin of Botany. 2026, 61(3):  402-415.  doi: 10.11983/CBB25087
Abstract ( 388 )   PDF (7168KB) ( 149 )   Save
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INTRODUCTION:  Glycosyltransferases are key enzymes involved in various biological processes in plants. To date, glycosyltransferases have been classified into 138 families, among which the glycosyltransferase family I is the largest. This family primarilyuses UDP-glucose as the glycosyl donor and is referred to as the UGT family. In recent years, the role of UGT genes in plant responses to abiotic stresses has gradually been revealed. However, currently, only a few UGT genes have been clearly identified as being involved in anthocyanin biosynthesis.


RATIONALE: To explore the mechanisms of anthocyanin and its glycosylation-related genes in tea plants under low-temperature stress, this study used FudingDabai Tea as the experimental material. Based on the transcriptome sequencing data, two genes, CsUGT73B4 (CSS0039619) and CsUGT85K5 (CSS0047548), which significantly upregulate anthocyanin expression, were identified. We further investigated the changes in these genes under low-temperature stress and explored the relationship between CsUGT73B4 and CsUGT85K5 genes and anthocyanin metabolism under low-temperature stress. Additionally, the functions of these two genes were verified through heterologous overexpression in Arabidopsis thaliana and gene silencing experiments.


RESULTS: Under low-temperature stress, the expression of CsUGT73B4 and CsUGT85K5 genes was positively correlated with anthocyanin accumulation. In Arabidopsis thaliana plants overexpressing these genes, anthocyanin content and antioxidant enzyme activities (SOD, POD, CAT) were significantly increased, while malondialdehyde (MDA) content and relative conductivity were reduced. Silencing of CsUGT85K5 led to feedback inhibition of CsUGT73B4 expression, revealing that these two genes act in concert to regulate the anthocyanin metabolic pathway.


CONCLUSION: In summary, the CsUGT73B4 and CsUGT85K5 genes are involved in the response to low-temperature stress, regulate anthocyanin metabolism, and may enhance the cold tolerance of tea plants.




The effect of low-temperature stress on anthocyanin in WTOE-CsUGT73B4 and OE-CsUGT85K5

(A) Observation of anthocyanin accumulation phenotype in Arabidopsis, the apical meristem position was marked with a white thread (The white line bar=1mm); (B) Effect of low-temperature stress on anthocyanin content in WT and OE-CsUGT73B4 Arabidopsis; (C) Effect of low-temperature stress on anthocyanin content in WT and OE-CsUGT85K5 Arabidopsis.  *P<0.05, **P<0.01, ***P<0.001.

The Mechanism of Manganese Accumulation Mediated by SpMTP10 Isolated from Sedum plumbizincicola
Siying Chen, Jinglin Wang, Yingyi Li, Xiangxin Lu, Peihong Zhang, Qinghong Qiu, Yan Gao, Tianyu Gu, Jiashi Peng
Chinese Bulletin of Botany. 2026, 61(3):  416-427.  doi: 10.11983/CBB25053   cstr: 32102.14.CBB25053
Abstract ( 243 )   PDF (4773KB) ( 900 )   Save
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INTRODUCTION: Manganese (Mn) is an essential micronutrient for plant growth and primarily act as enzyme cofactors and participate in the redox processes. However, excessive absorption of Mn by plant can also induce toxicity damages. Therefore, plants need to tightly regulate the uptake, homeostasis, and distribution of Mn to cope with stresses caused by its deficiency or excess. In these processes, the cation diffusion facilitator (CDF) family transporters, which in plants are also known as metal tolerance proteins (MTP), had been shown to be crucial for Mn homeostasis. Therefore, identifying MTP family genes and elucidating their underlying mechanisms for Mn accumulation would not only provide the novel insights about basic scientific issues of plant Mn accumulation, but also gene resources for crops improvement and Mn pollution bioremediation. 

RATIONALE: Sedum plumbizincicola is a recently discovered Cd/Zn hyperaccumulator that grows in mining areas. The soil in its natural habitat contains more than 10 000 mg·kg-1 of Mn, suggesting that S. plumbizincicola may have efficient Mn transport and detoxification capabilities. Based on the transcriptome sequencing results of S. plumbizincicola obtained previously, a member of the MTP family gene named SpMTP10 was cloned and its role in mediating Mn accumulation was investigated in this study. 

RESULTS: Phylogenetic analysis with orthologs from Arabidopsis and rice revealed that SpMTP10 belongs to the Mn-CDF subfamily and is most closely related to AtMTP10, AtMTP9 and OsMTP9, with the highest sequence identity of 72% to AtMTP10. SpMTP10 is mainly expressed in the roots of S. plumbizincicola and its expression level is not affected by Mn treatment. Expression of SpMTP10 in yeast can greatly enhance the tolerance of transformants to excessive Mn stress, and increase the Mn accumulation in transformants. However, under conditions of excessive cadmium (Cd), zinc (Zn), copper (Cu), and iron (Fe) stress, the yeast transformants exhibited no significant changes in tolerance. Subsequent subcellular localization analysis revealed that SpMTP10 was localized to the endoplasmic reticulum (ER) membrane. Compared with wild-type plants, transgenic Arabidopsis overexpressing SpMTP10 demonstrated reduced Mn accumulation in roots but increased Mn accumulation in shoots, rendering the plants more sensitive to excessive Mn stress. 

CONCLUSION: In conclusion, SpMTP10 likely enhances yeast tolerance to excessive Mn toxicity by promoting Mn sequestration in the ER. In plants, Mn transport mediated by SpMTP10 into the ER may facilitate intercellular migration of Mn in the ER lumen via plasmodesmata, thereby promoting Mn movement toward vascular tissues in roots and subsequent long-distance transport to shoots.

Analysis of the Function of WRI1 in Heat Stress of Arabidopsis Seedlings
Xuya Gu, Zhangman Lin, Siyuan Hu, Xuening Li, Xiaolin Qin, Zhengxi Wu, Nuo Li, Mingyin Feng, Ruihua Huang
Chinese Bulletin of Botany. 2026, 61(3):  428-436.  doi: 10.11983/CBB25085   cstr: 32102.14.CBB25085
Abstract ( 413 )   PDF (3542KB) ( 926 )   Save
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INTRODUCTION: Heat stress severely impairs plant growth and crop productivity. WRINKLED1 (WRI1), an AP2/EREBP-class transcription factor in Arabidopsis thaliana, orchestrates carbon partitioning between glycolysis and fatty acid biosynthesis, playing pivotal roles in development and stress adaptation. Elucidating its molecular function under high-temperature stress is critical for improving thermotolerance in crops. 

RATIONALE: While WRI1's metabolic regulatory function is established, its role in heat response remains unexplored. To decipher the molecular mechanism of WRI1-mediated thermotolerance, we integrated genetic approaches (wild-type, wri1-4, and WRI1-OE) with RT-qPCR and phenotyping under controlled heat stress. 

RESULTS:In this study, histochemical GUS staining of pWRI1::GUS transgenic lines demonstrated constitutive WRI1 expression throughout Arabidopsis seedlings, with significantly enhanced transcription in cotyledons under heat stress (HS) (P<0.05). Prolonged HS induced gradual transcriptional attenuation, though levels remained elevated versus optimal temperature (22°C). RT-qPCR confirmed thermo-responsive WRI1 upregulation (peak:1h HS, 3-fold induction), followed by threshold-dependent decline, indicating acute early-phase responsiveness. Endogenous immunoassays revealed reduced WRI1 protein accumulation under HS, suggesting HS-impaired protein stability or post-translational regulatory mechanisms. Thermotolerance phenotyping of WT, wri1-4, and WRI1-OE lines showed superior HS survival in WRI1-OE, with acquired thermotolerance exceeding basal thermotolerance across genotypes, confirming WRI1-mediated positive thermoregulation. The survival rate of WRI1-OE overexpression seedlings reached approximately 75%-85%, whereas that of wild-type and complementary lines was less than 10%. WRI1-OE reduced reactive oxygen species (ROS) accumulation, while RT-qPCR excluded direct transcriptional regulation of HSF/HSP genes (e.g., HSFA2, HSP101). Differential gene expression across genotypes nevertheless indicated WRI1's auxiliary role in thermotolerance via ROS scavenging and indirect proteostasis maintenance. 

CONCLUSION: This study establishes WRI1 as a master regulator of thermotolerance, functioning through synergistic activation of chaperone networks (HSFA2-HSPs) and reactive oxygen species (ROS) scavenging. The discovery of its cross-pathway coordination mechanism provides novel insights into plant thermal adaptation, while positioning WRI1 as an ideal target for breeding climate-resilient crops.

RESEARCH ARTICLE

Effect of Salt Stress on Carbohydrate Secretion by the Root System of Thinopyrum ponticum

Congcong Yi, Dan Li, Yanjie Li, Xiuwen Lin, Yingshuai Lu, Xiaopeng Chen
Chinese Bulletin of Botany. 2026, 61(3):  437-448.  doi: 10.11983/CBB25046   cstr: 32102.14.CBB25046
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INTRODUCTION: The carbohydrates secreted by plant roots play a crucial role in modulating the carbon source supply and metabolic activity of soil microorganisms.



RATIONALE: In order to unveil the response mechanism of root-secreted carbohydrates in salt-tolerant plants to the intensity of salt stress, this study focused on the salt-tolerant forage grass Thinopyrum ponticum. Four levels of salt stress intensity were conducted by adding NaCl, including control, light, moderate, and severe stress conditions (with soil salt contents of 0%, 0.2%, 0.4%, and 0.6% respectively). Untargeted metabolomics techniques were employed to analyze the compositional alterations of root-secreted carbohydrates and their correlations with the rhizosphere soil properties were evaluated.



RESULTS: The results demonstrated that: (1) Compared to the control, salt stress significantly decreased the contents of five types of carbohydrates, namely glycosides, aminoglycosides, D-glucosamine, 2-O-Methyl-L-fucose and galactoside, while markedly increasing glucose content; (2) Glycosides were the predominant carbohydrates secreted by Thinopyrum ponticum roots, accounting for 79.63% ± 8.19% of the total relative abundance, whereas the remaining five carbohydrate types ranged from 3.40% ± 0.53% to 4.57% ± 1.61%; (3) Root-secreted carbohydrates exhibited significant differences between the control and mild, moderate, or severe salt stress treatments, but no significant distinctions were observed among the latter three treatments; (4) Soil electrical conductivity and nitrate nitrogen content were identified as key factors responsible for the down-regulation of the five carbohydrate types, while soil pH was the primary factor driving the up-regulation of glucose.



CONCLUSION: Thinopyrum ponticum can adapt to salt-stressed environments by regulating the content of carbohydrates secreted by its roots. Meanwhile, the abundance of carbohydrates secreted by the roots is also influenced by the soil properties.




Conceptual map of the influence of salt stress on carbohydrate secretion by the roots of Thinopyrum ponticum



Genome-wide Identification of the AP2/ERF Gene Family and Its Expression Patterns under Salt Stress in the Desert Plant Reaumuria soongarica
Mengxuan Zhu, Yuan Liu, Haoyu Zhao, Zhenyun Sun, Wei Gong, Zhenhua Dang
Chinese Bulletin of Botany. 2026, 61(3):  449-461.  doi: 10.11983/CBB25066
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INTRODUCTION: Reaumuria soongorica, a perennial small shrub, is widely distributed in arid desert regions of northwestern China. It exhibits exceptional drought and salt tolerance, making it an ideal model for studying molecular mechanisms of plant stress resistance.


RATIONALE: We used the R. soongorica genome as a reference to identify members of the ‌AP2/ERF gene family‌ in this species, and ‌analyzed‌ their phylogeny, gene structure, conserved motifs, cis-acting elements, gene duplication events, ‌as well as‌ the expression patterns of these family members under salt stress.


RESULTS: ‌Seventy AP2/ERF genes‌ were identified from the R. soongorica genome. Phylogenetic analysis classified these genes into ‌four subfamilies: AP2, ERF, DREB, and RAV‌. Cis-acting element analysis revealed multiple regulatory elements associated with ‌light responsiveness, stress adaptation, growth regulation, and hormone signaling‌ in the promoter regions of R. soongorica AP2/ERF genes. These genes exhibited ‌an uneven distribution‌ across all 11 chromosomes of the R. soongorica genome, with ‌63 genes (90% of the total)‌ originating from gene duplication events. Evolutionary analysis suggested that ‌whole-genome duplication (WGD) and dispersed duplication‌ were the primary drivers of family expansion. Under salt stress, AP2/ERF genes showed ‌divergent expression patterns‌ in R. soongorica seedlings, with ‌six genes displaying significant differential expression‌ (|log2FC|≥1, P<0.05), implicating their potential roles in salt stress response.


CONCLUSION: This study identified and characterized the AP2/ERF gene family in R. soongorica at the genomic level, thereby establishing a foundation for elucidating its functional roles in this species' adaptation to arid and saline environments.




Phylogenetic tree of the AP2/ERF gene family in R. soongorica.


Comparative Analysis of Transcriptome of Adventitious Roots under Different Hydroponic Conditions of Lycium barbarum
Yang Gaier, Zhang Xuan, Wang Jiadong, Zhang Bo, Duan Linyuan, Li Xiang
Chinese Bulletin of Botany. 2026, 61(3):  462-474.  doi: 10.11983/CBB25077   cstr: 32102.14.CBB25077
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INTRODUCTION: This study investigates the transcriptional levels of adventitious root formation in different Lycium barbarum genotypes with varying root-forming abilities and aims to identify key genes involved in this process. The findings will provide a theoretical basis for in-depth research on the molecular mechanisms underlying adventitious root formation in wolfberry. 


RATIONALE: Three wolfberry genotypes with different root-forming abilities were used as experimental materials. A hydroponic experiment was conducted to analyze the transcriptional differences during adventitious root formation.


RESULTS: Transcriptome sequencing identified 6448 differentially expressed genes (DEGs), with the L-vs-H group having the highest number of DEGs at 4413, including 2583 upregulated and 1830 downregulated genes. A total of 281 transcription factors were identified, mainly from the MYB, AP2/ERF, and bHLH families, with distinct expression patterns. GO enrichment analysis revealed that 1714 DEGs were enriched in 32 GO terms. KEGG enrichment analysis indicated that DEGs were mainly enriched in the phenylpropanoid biosynthesis and plant hormone signal transduction pathways. Among these, MYB19 (Lba07g01820) is a core gene in the phenylpropanoid pathway, and TIR1 (Lba08g00069) is a core gene in the plant hormone signal transduction pathway. Both genes play crucial roles in adventitious root formation in wolfberry. qRT-PCR validated the reliability of the transcriptome data. 


CONCLUSION: This study elucidates the molecular mechanisms of adventitious root formation in wolfberry and lays a theoretical foundation for the genetic improvement and efficient propagation of wolfberry and other woody plants.



Schematic diagram of transcriptomic analysis of adventitic root generation under hydroponics of Lycium barbarum


TECHNIQUE AND METHOD
A Rapid In Planta Genetic Transformation System for MicroTom Tomato
Weikun Wang, Wenqian Fu, Qiumei Hong, Yang Zheng, Jiahui Yu, Yan Wang, Liangmiao Liu, Li Yang, Changchun Wang, Weidong Guo
Chinese Bulletin of Botany. 2026, 61(3):  475-484.  doi: 10.11983/CBB25061   cstr: 32102.14.CBB25061
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MicroTom tomato (Solanum lycopersicum cv. ‘MicroTom’) is widely used for functional characterization due to its short life cycle and clear genetic background. However, the traditional genetic transformation system of MicroTom based on tissue culture is constrained by low efficiency, long transformation period and complex operation. Based on the somatic cell reprogramming mechanism triggered by wound signaling, this study established an efficient and rapid in planta genetic transformation system for MicroTom tomato. Wound hypocotyl were created by removing apical cotyledons and true leaves from two-week-old seedlings, followed by direct inoculation with Agrobacterium tumefaciens carrying binary vector pCY-H05251-VcDAD2-EGFP (enhanced green fluorescent protein) to induce shoot regeneration. Results showed that a 28.6% PCR-based positive efficiency of regenerated shoots in the T0 generation, with seeds derived within 4–5 months post inoculation. Antibiotic and fluorescence screening revealed approximate 73.5% lines in the T1-generation expressed the fused EGFP protein. Compared to conventional tissue culture-dependent transformation systems, this protocol enhanced transformation efficiency, shortened transformation period, and simplified sterile operational procedures. The in planta genetic transformation system provides a robust platform for functional genomics studies, and significantly lowers technical barriers in tomato genetic breeding.
Establishment of Regeneration System in vitro for Hydrangea macrophylla cv. ‘Chikushi-no-kaze’
Yuyan Jin, Shuangshuang Chen, Jing Feng, Xintong Liu, Xiangyu Qi, Huijie Chen, Yan Dong, Yanming Deng
Chinese Bulletin of Botany. 2026, 61(3):  485-495.  doi: 10.11983/CBB25068   cstr: 32102.14.CBB25068
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INTRODUCTION: Hydrangea is essential in landscaping, ecology, and medical care, with significant development prospects. ‘Chikushi-no-kaze’ is an ideal low-maintenance variety for micro-landscaping and potted Hydrangea, widely favored by consumers. However, the extremely low setting rate of Hydrangea and poor seed development under natural conditions render traditional reproduction methods inadequate for meeting the demands of large-scale annual production in the market. The breeding of Hydrangea plantlets through tissue culture technology is currently the most efficient method for producing high-quality plantlets. 

RATIONALE: Regeneration efficiency in plant tissue culture is a key factor in achieving factory seedling production. Therefore, this study investigated the regeneration efficiency of isolated leaves from tissue culture plantlets of ‘Chikushi-no-kaze’ under optimal culture conditions at each key stage, considering different leaf positions, dark culture durations, and other factors. The aim was to establish an efficient regeneration technology system, which provides technical guidance for large-scale plantlet production and serves as a reference for establishing a genetic transformation system for Hydrangea in the future. 

RESULTS: The 3rd to 5th leaves (middle mature leaves) of ‘Chikushi-no-kaze’ tissue culture plantlets were identified as the optimal sampling leaves. A dark culture duration of 10-14 days was conducive to callus formation. The most suitable medium for the induction and regeneration of adventitious buds was MS + 3.0 mg·L-1 CPPU + 0.1 mg·L-12,4-D, with induction and regeneration rates of 97.78% and 93.33%, respectively. The optimal medium for adventitious bud proliferation was MS + 2.0 mg·L-1 6-BA + 0.1 mg·L-1 IBA, yielding a proliferation coefficient of 8.33. For elongation growth, the optimal medium was MS + 1.0 mg·L-1 6-BA + 0.1 mg·L-1 IBA, resulting in an average stem length of 4.10 cm. The optimal medium for rooting culture was MS + 0.3 mg·L-1 IBA, achieving a rooting rate of 87.20%. 

CONCLUSION: This study initially established a technical system for in vitro leaf regeneration of big leaf Hydrangea 'Chikushi-no-kaze', which effectively solved the problem of low efficiency of adventitious bud regeneration of Hydrangea, helped to achieve efficient reproduction and recycling. Lay the foundation for large-scale production and genetic improvement of Hydrangea.

Establishment of a Regeneration System for Changnienia amoena
Shengfei Yang, Yuye Deng, Shiyun Cai, Yafei Liu, Yin Peng, Yuanjie Ding
Chinese Bulletin of Botany. 2026, 61(3):  496-505.  doi: 10.11983/CBB25081   cstr: 32102.14.CBB25081
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INTRODUCTION: Changnienia amoena is listed as a national second-class protected plant. It is a rare orchid species unique to China, as well as a valuable medicinal plant and a potential wild potted flower germplasm resource with great development potential. Currently, wild resources of C. amoena are rapidly declining, making conservation efforts crucial. 

RATIONALE: Research on C. amoena regeneration technology has mainly focused on different combinations of plant growth regulators, and a complete regeneration system has not yet been established. Tissue culture technology plays a crucial role in the conservation of wild C. amoena resources, and research on its regeneration system can promote the sustainable development and utilization of these resources. 

RESULTS: The results showed that: (1) 75% ethanol had a significant effect on the sterilization of C. amoena pseudobulbs, and the best sterilization treatment for C. amoena pseudobulbs was to use 75% ethanol for 30 seconds, followed by 0.1% mercuric acid solution for 12 minutes; (2) The pseudobulbs were able to induce adventitious buds under the different blocking treatments, but the effect of the adventitious buds was that the complete pseudobulbs> the halved pseudobulbs> the cruciform block pseudobulbs and the rate of germination was much higher than that of the blocking treatment; (3) Considering the browning rate, contamination rate, survival rate and induced germination rate, the optimal collection time for C. amoena pseudobulbs to be used as explants for adventitious shoots induced in May; (4) The optimal medium for the adventitious shoots of C. amoena pseudobulbs was 1/2 MS+1.0mg∙L–1 6-BA+ 0.5mg∙L–1 NAA, with an induction rate of 75.56%; the optimal rooting medium is1/2 MS+0.4 mg∙L–1 6-BA+1.0 mg∙L–1 NAA, with a rooting rate of up to 93.33%; (5) After treatment with a 50 mg∙L–1 6-BA solution, the histocultured plantlets were transplanted into humus soil and covered with plastic film for hardening. The survival rate was as high as 94.44%. 

CONCLUSION: In this study, the pseudobulbs of C. amoena were used as explants to explore the effects of sterilization conditions, the way of explant cutting, different collection times, and the concentration of plant growth regulators on the sterilization effect of explants, inducing adventitious shoots and rooting of histocultured plantlets, and to conduct a preliminary study on the regeneration system of C. amoena, which can help to protect the germplasm resources of C. amoena, and provide a theoretical basis and technical references for the future propagation of C. amoena.

TECHNIQUE AND METHOD
Establishment of a Universal Tissue Culture System and Anatomical Structure Characteristics of Succulent Plants
Mengqi Wen, Yinghong Tang, Cancai Zhao, Min Wu, Yiqian Zheng, Lu Luo, Jianrong Chen
Chinese Bulletin of Botany. 2026, 61(3):  506-518.  doi: 10.11983/CBB25154   cstr: 32102.14.CBB25154
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INTRODUCTION: Succulent plants have morphological diversity, high added value, and strong stress resistance. However, the industrialization of these plants encounters obstacles such as the challenge of isolating propagation traits, sluggish proliferation rates, marked interspecific variations, and the intricacies involved in generalizing tissue culture parameters across different groups. 




RATIONALE: Therefore, the three succulent plants (Echeveria×Sedeveria 'Pink Rubby', Crassula fusca, and Aporocactus flagelliformis) from two families and three genera were selected as experimental materials in this study. The investigation of the universal tissue culture system has been conducted, including procedures for disinfecting leaf/stem explants, differentiating adventitious buds, rooting adventitious buds (using traditional and open tissue culture methods), and transplanting. The anatomical structure characteristics of leaf/stem in tissue culture plantlets and potted seedlings were analyzed by frozen section method. 


RESULTS: The result revealed that the optimal medium for leaf/stem differentiation adventitious bud was MS supplemented with 3.0 mg·L-1 6-BA and 0.8 mg·L-1 NAA, achieving an induction rate of over 95%. For adventitious bud rooting, the optimal medium of MS with 4.0 mg·L-1 IBA, 2.0 mg·L-1 NAA, and 0.4 mg·L-1 KT, further enhanced with an additional 0.3 mg·L-1 S206 in open tissue culture, resulted in a rooting rate exceeding 90%. The optimal universal transplanting substrate was a mix of grass charcoal soil and sand at a ratio of 2:1 by volume, resulting in a survival rate exceeding 99%. In addition, the epidermal thickness of tissue cultured plantlets of three succulent plants were significantly thinner than that of wild seedlings. Only the vascular bundle area of Aporocactus flagelliformis was much smaller than that of wild seedlings, but the number of vascular bundles of tissue cultured plantlets was significantly higher than that of wild seedlings.








 

  

Tissue culture plantlet and their anatomical structure of three succulent plants.


 

CONCLUSION: These findings offer theoretical underpinning and practical guidance for establishing an efficient cross-group propagation system leveraging physiological structural traits, advancing cost-effective pollution prevention and control solutions, and facilitating industrial advancement.


SPECIAL TOPICS
Research Progress on Effectors of Arbuscular Mycorrhizal Fungi
Yuexuan Zhang, Peng Wang
Chinese Bulletin of Botany. 2026, 61(3):  519-528.  doi: 10.11983/CBB25079   cstr: 32102.14.CBB25079
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Arbuscular mycorrhizal fungi (AMF) can form symbiotic relationships with approximately 80% of terrestrial plants. Through their unique arbuscular structures within roots, they establish close contact with host cells to create a bidirectional nutrient exchange interface. This mutualistic mechanism not only enhances plant stress resistance but also reshapes ecosystem nutrient cycling. Like pathogenic fungi, the cell walls of AMF are primarily composed of chitin and β-glucans, which are key molecular patterns capable of triggering host plant immune responses. How AMF effectively evades host plant immunity remains unclear. Effector proteins secreted by pathogenic fungi have been found to play a crucial role in suppressing plant immune responses. During arbuscular mycorrhizal symbiosis, numerous effector proteins are also induced, which may similarly inhibit plant immunity and facilitate fungal colonization. This article reviewed and summarized current research on AMF effector proteins and discussed the future research directions and challenges. Studying effector proteins will help elucidate the regulatory mechanisms underlying the establishment and maintenance of AMF symbiosis, deepen our understanding of host-fungal interactions, and aid in selecting optimal fungal strains and plant varieties for enhanced symbiotic efficiency, thereby promoting sustainable agricultural development.
Research Progress on the Induction of Leaf Color Mutants in Medicinal Plants and Their Molecular Mechanisms of Mutation
Shengyun Wang, Erbiao Guo, Mengjun Li, Zhonghua Wang
Chinese Bulletin of Botany. 2026, 61(3):  529-540.  doi: 10.11983/CBB25069   cstr: 32102.14.CBB25069
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This review focuses on leaf color mutants of medicinal plants, systematically expounds their induction pathways, mutation molecular mechanisms and characteristic applications, and highlights the research value of medicinal plants in secondary metabolism regulation. Leaf color mutants are mutation types in which gene mutations cause abnormalities in chlorophyll synthesis or degradation, thereby changing leaf color. They can be classified according to seedling leaf color, pigment content, genetic characteristics, etc. Their induction methods are divided into spontaneous mutation and artificial induction mutation, and the latter covers physical, chemical and biological mutagenesis, each with its own advantages and disadvantages. In terms of molecular mechanisms, mutations in key genes for chlorophyll synthesis and degradation lead to pigment metabolism imbalance, abnormalities in chloroplast development genes affect chloroplast structure and function, variations in photosynthesis genes change the efficiency of light energy capture and conversion, and transcription factors and light signal/hormone pathways synergistically regulate leaf color. Especially in medicinal plants, leaf color mutations are often accompanied by changes in photosynthetic efficiency. Through energy supply, carbon-nitrogen allocation and metabolic precursor sharing, the "chlorophyll metabolism-secondary metabolism" network is reshaped to regulate the synthesis and accumulation of medicinal secondary metabolites such as flavonoids, terpenoids and alkaloids. Therefore, leaf color mutants are not only tools for analyzing photosynthetic and chloroplast development mechanisms, but also key materials for mining the regulatory network of medicinal component synthesis, and have broad application prospects in functional genomics research, molecular marker-assisted breeding and the creation of medicinal germplasm with high active ingredients. Although current research has achieved certain results, it still faces problems such as low efficiency in mutant screening, unclear functions of some genes, and insufficient integration of multi-omics data. In the future, relying on technological innovations such as CRISPR gene editing, combining multi-omics integration and artificial intelligence screening, we should focus on breaking through the light regulation mechanism of medicinal component synthesis and promoting the genetic improvement of endangered species and the cultivation of high-active varieties.
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