Jiaxia Zhang

Adventitious root (AR) formation is the key step for successful cutting propagation of tea plants (Camellia sinensis L.). Studies showed that arbuscular mycorrhizal fungus (AMF) can promote the rooting ability, and auxin pathway in basal stem of cuttings was involved in this process. However, auxin and abscisic acid (another important regulator on AR formation) in the other parts of cuttings at different rooting stages responding to AMF inoculation are not well studied. Therefore, in this paper, contents, enzymes and genes related to these two plant hormones were comprehensively determined aiming to unveil how endogenous indole-3-acetic acid (IAA) and abscisic acid (ABA) involve in the AMF regulating AR development of tea cuttings. Inoculating with AMF significantly increased the proportion of cuttings at S2 stage (AR formation), which was more than twice as much as the control. And the total rooting rate in mycorrhizal treatment was also higher than that in the control with an increase of 8.66%. Enzyme activity assays showed that except for decreased polyphenol oxidase (PPO) activity at the S3 stage and peroxidase (POD) activity in middle stem of S3 stage, AMF inoculation increased activities of POD, PPO, superoxide dismutase (SOD) and catalase (CAT) to varying degrees in leaf, middle stem and basal stem of tea cuttings. After inoculation with AMF, the indoleacetic acid oxidase (IAAO) activity decreased to a certain extent in the first three stages of tea cuttings, which showed a trend of ‘low-high-low’ in the basal stem of all treatments. Besides, there was a significantly positive correlation between SOD activity and AR formation, especially for the proportion of cuttings at S2 and S3 stages. Higher IAA level and IAA/ABA ratio was found in basal stem of cuttings at S1 stage induced by AMF, which promoting the AR formation as revealed by correlation analysis. At the same time, AMF significantly elevated the level of IAA in leaf at S1 stage. By screening differentially expressed genes (DEGs) related to IAA and ABA pathways, together with redundant analysis, it was indicated that auxin biosynthesis and transport, as well as ABA transport and signal transduction, were involved in AMF regulating the rooting of tea cuttings. Overall, both endogenous IAA and ABA played roles in the regulation of AR formation of tea cuttings by AMF inoculating, enriching the theoretical basis of AMF regulating rooting of cuttings and providing foundations for cutting propagation of tea plants.

Introduction: Adventitious root (AR) development, affected by various biotic and abiotic factors, is the most important procedure in tea plant (Camellia sinensis L.) cutting propagation. Establishing symbiotic relationships with most terrestrial plants, AMF (Arbuscular mycorrhizal fungus) can mediate the AR formation of several herbaceous and woody plants in previous studies.Methods: In this paper, effects of combined application of AMF and exogenous auxin on AR formation of cuttings from different tea plant varieties (‘Pingyangtezao’, ‘Longjing 43’ and ‘Longjingchangye’) were studied. Then we also performed RNA-Seq analysis with ‘Pingyangtezao’ cuttings aiming to find the possible auxin-related pathway of AM fungal regulation on AR formation. To accurately uncover the regulatory mechanism of AMF on AR formation of tea cuttings, rooting process were separated into four stages (S0, non-rooting; S1, AR protrusion; S2, AR formation and S3, AR elongation) at the same sampling time.Results and Discussion: Results showed that IBA treatment increased the mycorrhizal colonization rate, especially in ‘Pingyangtezao’ variety (from 37.58% to 46.29%). Both inoculating AMF and addition of IBA promoted the AR formation, and rooting of different tea plant varieties showed different dependence on auxin. AMF could alleviate the effect of auxin-related inhibitors (2,3,5-triiodobenzoic acid, L-α-(Aminooxy)-β-phenylpropionic acid and α-(phenylethyl-2-oxo)-IAA) on rooting of tea cuttings, even though the colonization of AMF was hindered at various degrees. Transcriptomic analysis showed that different numbers of differentially expressed genes (DEGs) at various rooting stages of tea cuttings with the most at S2 stage (1360 DEGs), indicating the increasing regulation by AMF with the development of AR. Similar trend was found in auxin-related DEGs, and family genes of YUC, GH, PIN, LAX, SAUR, AUX, and ABP involved in the AM fungal regulation on AR formation of tea cuttings. Additionally, AMF strongly mediated auxin transport and signal transduction pathways in tea cuttings as showed by the results of correlation analysis. Overall, interaction of AMF and exogenous auxin in promoting rooting and the preliminary mechanism of AMF regulating AR formation of tea cuttings was deciphered in this paper, which may provide a basis for further deep mechanistic research and cutting propagation of tea production.

Tea has been gaining increasing popularity all over the world in recent years, and its yield and quality depend on the growth and development of tea plants [Camellia sinensis (L.) Kuntze] in various environments. Nowadays, biotic stress and extreme weather, such as high temperature, drought, waterlogging, pests, and diseases, bring about much pressure on the production of tea with high quality. Wherein anthracnose, which is the most common and serious disease of tea plants, has earned more and more attention, as its control mainly relies on chemical pesticides. Arbuscular mycorrhizal fungi (AMF), forming symbiosis with most terrestrial plants, participate in plant resistance against the anthracnose disease, which was found by previous studies in a few herbaceous plants. However, there are a few studies about arbuscular mycorrhizal (AM) fungal regulation of the resistance to the anthracnose pathogen in woody plants so far. In this paper, we investigated the effect of AMF on the development of anthracnose caused by Colletotrichum camelliae and tried to decipher the pertinent mechanism through transcriptome analysis. Results showed that inoculating AMF significantly reduced the damage of anthracnose on tea seedlings by reducing the lesion area by 35.29% compared to that of the control. The content of superoxide anion and activities of catalase and peroxidase significantly increased (P < 0.05) in mycorrhizal treatment in response to the pathogen with 1.23, 2.00, and 1.39 times higher, respectively, than those in the control. Pathways of plant hormone signal transduction, mitogen-activated protein kinase (MAPK) signaling, and phenylpropanoid biosynthesis might play roles in this regulation according to the transcriptomic results. Further redundancy analysis (RDA) and partial least squares structural equation modeling (PLS-SEM) analysis found that plant hormones, such as auxin and ethylene, and the antioxidant system (especially peroxidase) were of great importance in the AM fungal alleviation of anthracnose. Our results preliminarily indicated the mechanisms of enhanced resistance in mycorrhizal tea seedlings to the anthracnose pathogen and provided a theoretical foundation for the application of AMF as one of the biological control methods in tea plantations.

Arbuscular mycorrhizal fungus (AMF), forming symbiosis with most terrestrial plants, strongly modulates root system architecture (RSA), which is the main characteristic of root in soil, to improve plant growth and development. So far, the studies of AMF on tea plant seedlings are few and the relevant molecular mechanism is not deciphered. In this study, the 6-month-old cutting seedlings of tea plant cultivar “Wancha No.4” were inoculated with an AMF isolate, Rhizophagus intraradices BGC JX04B and harvested after 6 months of growth. The indexes of RSA and sugar contents in root were determined. The transcriptome data in root tips of mycorrhizal and non-mycorrhizal cutting seedlings were obtained by RNA-sequence (Seq) analysis. The results showed that AMF significantly decreased plant growth, but increased the sucrose content in root and the higher classes of lateral root (LR) formation (third and fourth LR). We identified 2047 differentially expressed genes (DEGs) based on the transcriptome data, and DEGs involved in metabolisms of phosphorus (42 DEGs), sugar (39), lipid (67), and plant hormones (39) were excavated out. Variation partitioning analysis showed all these four categories modulated the RSA. In phosphorus (P) metabolism, the phosphate transport and release (DEGs related to purple acid phosphatase) were promoted by AMF inoculation, while DEGs of sugar transport protein in sugar metabolism were downregulated. Lipid metabolism might not be responsible for root branching but for AMF propagation. With respect to phytohormones, DEGs of auxin (13), ethylene (14), and abscisic acid (5) were extensively affected by AMF inoculation, especially for auxin and ethylene. The further partial least squares structural equation modeling analysis indicated that pathways of P metabolism and auxin, as well as the direct way of AMF inoculation, were of the most important in AMF promoting root branching, while ethylene performed a negative role. Overall, our data revealed the alterations of genome-wide gene expression in tea plant roots after inoculation with AMF and provided a molecular basis for the regulatory mechanism of RSA (mainly root branching) changes induced by AMF.

Being one of the most disastrous pierce-sucking pests of tea plants [Camellia sinensis (L.) O. Kuntze], tea green leafhopper (Empoasca onukii Matsuda) greatly affects tea yield and quality. In this paper, correlations among leaf structure, biochemical components and tea green leafhopper occurrence in tea cultivars with larger planting area were deeply analyzed, aiming to find the key factor impacting the host selectivity of tea green leafhopper. Tea cultivar ‘Longjing 43’, together with ‘Shuchazao’, showed the most susceptibility to this pest with high tea polyphenols content. And the best resistant tea plant cultivar, ‘Caoxi No.1’, largely relied on the thicker leaf, epidermis (both upper and lower), cuticle and spongy tissue as revealed by principal component analysis. Here, we suggested that tea polyphenols were considered as a biochemical marker for determining the resistance ability to tea green leafhopper of different tea cultivars, along with leaf upper and lower structures as mechanical barriers, which can potentially be used to promote breeding of insect-resistant tea plants.