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DoSPX1 and DoMYB37 regulate the expression of DoCSLA6 in Dendrobium officinale during phosphorus starvation

BMC Plant Biology volume 24, Article number: 803 (2024) Cite this article

Abstract

Background

Dendrobium officinale Kimura et Migo (D. officinale) is parasitic on rocks or plants with very few mineral elements that can be absorbed directly, so its growth and development are affected by nutritional deficiencies. Previous studies found that phosphorus deficiency promotes polysaccharides accumulation in D. officinale, the expression of DoCSLA6 (glucomannan synthase gene) was positively correlated with polysaccharide synthesis. However, the molecular mechanism by which the low phosphorus environment affects polysaccharide accumulation remains unclear.

Results

We found that DoSPX1 can reduce phosphate accumulation in plants and promote the expression of PSIs genes, thereby enhancing plant tolerance to low phosphorus environments.Y1H and EMSA experimental show that DoMYB37 can bind the promoter of DoCSLA6. DoSPX1 interact with DoMYB37 transiently overexpressed DoSPX1 and DoMYB37 in D. officinale protocorm-like bodies, decreased the Pi content, while increased the expression of DoCSLA6.

Conclusions

The signaling pathway of DoSPX1-DoMYB37-DoCSLA6 was revealed. This provides a theoretical basis for the accumulation of polysaccharide content in D. officinale under phosphorus starvation.

Peer Review reports

Background

D. officinale which is the second largest genus in the orchid family is a perennial epiphytic herb. D. officinale has a variety of pharmacological effects including antioxidant and immune-enhancing properties [1,2,3]. Mannose and glucose were the most abundant monosaccharide component in soluble polysaccharides, and were the most important component affecting the efficacy of soluble polysaccharides [4, 5]. The total polysaccharide and mannose content are quantitative indicators of D. officinale [6].

Phosphorus (Pi) plays an important role in various physiological processes in plants. The majority of the phosphorus in the soil forms adsorbed states, resulting in long-term phosphorus deficiency in plants [7,8,9]. D. officinale is generally attached to large tree trunks in primitive forests, shady and damp cliffs, and rock crevices. It is important to study how D. officinale ironwood absorbs and utilizes phosphate and accumulates polysaccharides in this growth environment. Our previous study founded that inorganic phosphate (Pi) deficiency promoted the accumulation of D. officinale polysaccharides [10]. Under LP environments, the expression of DoCSLA6, a member of the CSLA family directly involved in mannose synthesis, was highly positively correlated with the polysaccharide accumulation [10, 11]. However, the molecular mechanism of polysaccharide accumulation induced by low phosphorus in D. officinale remains unclear.

The SPX domain was coined based on the first three letters of Suppressor of Yeast gpa1 (SYG1), Phosphatase 81 (PHO81) and Xenotropic and Polytropic Retrovirus receptor1 (XPR1). Proteins with the SPX domain have been identified as the most important regulators of plant responses to LP, SPX usually works with downstream transcription factors [12,13,14,15]. In soybean, GmSPX1 is a negative regulator in response to phosphorus signaling. GmMYB48 is a phosphate starvation-inducible transcription factor. GmSPX1 overexpression decreased the transcripts of AtMYB4, an ortholog of GmMYB48. [16]. In Arabidopsis thaliana, AtSPX4 can interact with AtPAP1 to regulate anthocyanin synthesis in a phosphate-dependent manner [17]. Phylogenetic analysis suggests that DoSPX1 may be involved in LP signaling pathway in D. officinale [18].

In this study, we analyze the function of DoSPX1 and DoMYB37 by overexpression of DoSPX1 and DoMYB37 in Nicotiana tabacum L.(N. tabacum) and D. officinale protocorm-like bodies. We confirmed the interaction between DoSPX1 and DoMYB37 by Yeast Two-Hybrid Assay (Y2H) and Bimolecular fluorescence complementation (BIFC). We also verified the transcriptional regulation of DoCSLA6 by DoMYB37.

Materials and methods

Plant materials and growth conditions

D. officinale were selected from the Engineering Technology Research Center for the Development and Utilization of Local Characteristic Plant Resources in Anhui Province, Anhui Agricultural University. The D. officinale was cultured in MS medium with a light period of 16 h and incubation temperature of 25 degrees Celsius.

Nicotiana tabacum and Yunyan seedlings were cultured on Murashige and Skoog (MS) medium at 25 °C, which were lighted for 16 h and dark for 8 h.

Plants were cultured under phosphorus concentrations of 0.0625 mM (LP) and 1.25 mM (NP) master mixes (Supplementary Table 1). MS media with different phosphorus concentrations were prepared using the same molar concentration of KCl instead of KH2PO4.

Quantitative real-time PCR (qRT-PCR)

The RNA was extracted from the Liquid nitrogen quick-frozen plant tissues using the Plant Total RNA Isolation Kit. A One Step RT-qPCR Kit was used to obtain cDNA. 2 × TaqMan Fast qPCR Master Mix was used to execute qRT-PCR. Reaction conditions were performed according to Liu’s method [10]. The qRT-PCR primers were designed using NCBI PRIMER-BLAST8 (Supplementary Table 2). The relative quantitative analysis method (2−ΔΔCT) was used to calculate the relative gene expression, and all experiments were repeated with more than three biological replicates.

Subcellular localization analysis

To analyze the subcellular localization of the DoSPX1 and DoMYB37 protein, the open reading frame (ORF) was amplified by PCR from the D. officinale cDNA library with DoSPX1 (LOC110102021) and DoMYB37 (LOC110099272) gene-specific primers (Supplementary Table 2) and inserted into the plant expression vector pCAMBIA1305.1-GFP to generate the pCAMBIA1305.1-DoSPX1 and pCAMBIA1305.1-DoMYB37 construct. Cells of the GV3101-pSoup-p19 carrying pCAMBIA1305.1-DoSPX1/DoMYB37 and pCAMBIA1305.1-GFP (positive control) were separately infiltrated into Nicotiana tabacum. Transient expression of GFP signals were observed using a laser confocal microscope after 2 d of infiltration.

Yeast two hybrid (Y2H)

The full-length ORF of DoSPX1 and DoMYB37 were constructed on pGADT7 and pGBKT7 respectively. Recombinant vector pGADT7-DoSPX1 and pGBKT7-DoMYB37 were expressed in AH109 strains. Then transformed clones were screened on SD-WL (SD medium without Leu and Trp), and clones were screened on SD-HAWL (SD medium without Ade, His, Leu, and Trp). X-α-Gal is used to identify positive interactions.

Bimolecular fluorescent complementary assay (BiFC)

The full-length ORF of DoSPX1 and DoMYB37 were constructed on pCAMBIA2300s-YC and pCAMBIA1300s-YN respectively. Single colonies of Agrobacterium were picked and propagated in YEP medium and incubated in a shaker at 28 °C for 24 h. The bacterial sap was collected by centrifugation and resuspended to OD600 = 0.2 using onion infestation solution, fresh red onions were taken, the epidermis was removed from the third or inner scale, immersed in the onion infestation solution, and incubated in the dark for 30 min. After the infestation, the excess was aspirated from the epidermis using filter paper and incubated for 20–24 h at 22 °C in the dark, and the fluorescence signal was observed using a laser confocal microscope.

Yeast one-hybrid assay (Y1H)

The DoCSLA6 promoter cis-acting elements were predicted using the online tools PlantCARE (http://bioinformatics.psb.ugent.be/webtools/plantcare/html/) and New PLACE (https://www.dna.affrc.go.jp/PLACE/?action=newplace) [19].

The gene promoter is expressed at the background level in Saccharomyces cerevisiae Y1Hgold and the minimum/critical ABA concentration for growth inhibition is determined by adding different concentrations of ABA to inhibit the interaction of the promoter fragment itself. The complete DoMYB37 ORF sequence was constructed on the pGADT7 vector to form a recombinant vector, and the MBS in the promoters of DoCSLA6 of glucomannan biosynthesis were constructed on the pABAi vector. The DoMYB37 recombinant plasmids were expanded and cultured with the pGADT7 vector attached and the plasmids were extracted for use. The yeast strain containing the target promoter was prepared as receptor cells and the transformed strain was coated onto SD/-Leu plates with appropriate concentrations of ABA and incubated at 28 °C for 2–3 d. Then the positive colony was picked and tested for growth of monoclonal strains on SD/-Leu medium for 48 h. Empty vectors pGADT7 were used as negative controls.

Electrophoretic mobility shift assay (EMSA)

The target gene promoter sequences were analyzed through the NEWPLACE online (https://www.dna.affrc.go.jp/PLACE/?action=newplace) and the promoter probes were designed. The ORF of DoMYB37 was inserted into the vector pGEX4T-1. Recombinant protein DoMYB37-GST was expressed in E. coli BL21 (DE3) and purified using a protein purification kit with a GST-tag. Probes were mixed in equal proportions upstream and downstream, denatured at 100 °C for 10 min, and then slowly brought to room temperature and set aside. DoMYB37-GST purified protein was combined with probes of proDoCSLA6 by adding 5 × binding buffer at 25 °C for 25 min. Then 6 × Loading Buffer was added to the system and the sample was loaded and electrophoresed at 120 V for 45 min. After electrophoresis, the PAGE gel was removed and stained with nucleic acid dye for 15 min and the results were observed using a gel imager (Shanghai Tanon).

N. tabacum conversion methods

To analyze the DoSPX1 and DoMYB37, The ORF of DoSPX1 and DoMYB37 was inserted into the pCAMBIA1305.1 vector. Agrobacterium tumefaciens was expanded and collected and resuspended in infestation solution to OD600 = 0.4–0.6 and set aside. N. tabacum with 2–4 true leaves were cut out and placed in dark culture in MS budding medium for 3 days. The dark-cultured leaves were placed in the infestation solution, blotted on filter paper to dry the surface of the infestation solution, and raised in solid medium containing acetosyringone for 3 d. The leaves were washed 2–3 times with sterile water containing Temetin, blotted on filter paper to dry the surface liquid, placed in MS germination medium and incubated at 28 °C in the light, and decolonized again within two weeks. N. tabacum shoots up to 1 cm in length were placed in MS empty medium for rooting, and after rooting, the seedlings were transplanted.

Transformation of D. officinale protocorm-like bodies

In this laboratory, seeds were used as explants to induce D. officinale protocorm-like bodies, and the experimental materials were preserved by the laboratory, and their traits were genetically stable. D. officinale protocorm-like bodies were incubated in the dark for 3 days and the Agrobacterium introduced into the plasmid was expanded and cultured. Bacteria were collected and resuspended in an infestation solution to OD600 = 0.4–0.6. After washing 2–3 times in sterile water containing antibiotics, the surface was dried with filter paper and then placed in a decontamination medium for sterilization and screening (Supplementary Fig. 1).

Determine the Soluble phosphorus content of plants

Determination of soluble Pi content of plant materials by ammonium molybdate method. We grind the plants with liquid nitrogen and 10% sodium perchlorate, then dilute the supernatant tenfold with 5% (w/v) perchloric acid, centrifuge, and then tenfold with 5% (w/v) perchloric acid. Working solution [ammonium sulfate-molybdate (solution A) and ascorbic acid (solution B) mixed in a ratio of (6:1)]. Calculate the phosphorus content by measuring the absorbance at 820 nm using a UV spectrophotometer.

Results

Overexpression of DoSPX1 in N. tabacum

We first verified the subcellular localization of DoSPX1, the ORF of DoSPX1 was inserted into the pCAMBIA1305.1 vector. The results showed that the fusion protein DoSPX1-GFP was mainly distributed in the nucleus (Supplementary Fig. 2). At the same time, we constructed tobacco overexpression lines of DoSPX1. Compared to N. tabacum transferred with pCAMBIA1305 empty vector, DoSPX1 transgenic N. tabacum showed a significant reduction in leaf number and a significant increase in root length and Root to Shoot ratio, 1.34 and 3.82 times higher than the control, respectively (Fig. 1A, B). The Pi content of stems and roots in transgenic N. tabacum decreased under low phosphorus treatment compared to the control (Fig. 1C). The relative expression of DoSPX1 was significantly increased by phosphorus starvation, and the changes were more pronounced in the roots (Fig. 1D). When plants are stressed by LP, a large number of Pi starvation-induced (PSI) genes, such as phosphorus starvation response(PHRs), phosphate transporters(PTs) are induced. In the OE-DoSPX1 transgenic lines,PSI genes were promoted more obviously by LP (Fig. 1E). These studies suggest that DoSPX1 overexpression can reduce phosphate accumulation in plants and promote the expression of PSIs genes, thereby enhancing plant tolerance to LP environments.

Fig. 1
figure 1

Phenotype and gene expression in OE-DoSPX1 transgenic N. tabacum and emptor vector (control group). A OE-DoSPX1 transgenic N. tabacum (The right) and control group (The left). B Phenotypic (number of blades, root length, root-shoot ratio) differences in OE-DoSPX1 N. tabacum and control group. C Soluble phosphorus content in OE-DoSPX1 N. tabacum and control group. D Expression of DoSPX1 in OE-DoSPX1 N. tabacum and control group. E Expression of PSI genes (NtPHR1, NtPHR2, NtPT1, NtPT2) in OE-DoSPX1 N. tabacum and control group. Significant differences between the Empty vector and OE-DoSPX1 seedlings grown under the same conditions (HP and LP) are indicated using asterisks (**p < 0.01; *p < 0.05; two-tailed Student’s t-test)

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DoMYB37 responds to LP environment and interacts with DoSPX1

Many other researchers have previously found that the 20th subfamily R2R3-MYB are response to phosphate deficency. Phylogenetic evolutionary showed that DoMYB37 belongs to the 20th subfamily of the MYB family. The expression levels of DoMYB37 were increased with decreasing phosphorus concentration in D. Officinale (Supplementary Fig. 3).

DoMYB37-GFP are subcellular localized to the nuclear (Fig. 2A). To identify the interaction between DoMYB37 and DoSPX1, we divide DoMYB37 into N and C terminals. (Fig. 2B). The Y2H showed that DoMYB37-N grew with the yeast strain of DoSPX1 and showed blue color, indicated that DoSPX1 interacted with the N-terminal of DoMYB37 (Fig. 2C). BiFC verified the interaction of DoMYB37 and DoSPX1 in onion cells, and the YFP signaling is localized in the nucleus of onion cells (Fig. 2D).

Fig. 2
figure 2

Subcellular localization of DoMYB37 and interaction between DoMYB37 and DoSPX1. A Subcellular localization of DoMYB37. B Structural domain analysis and truncation of DoMYB37 proteins. C Yeast two-hybridization of DoMYB37-N with DoSPX1. D BIFC analysis (YFP: Yellow fluorescent signals)

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Functional identification of DoMYB37 in N. tabacum and D. officinale

Transgenic N. tabacum. overexpressing DoMYB37 had shorter stems and longer root systems compared with the control group (Fig. 3A). OE-DoMYB37 N. tabacum plants had 1.27 times longer root length compared to the empty vector (Fig. 3B). The expression levels of NtPT1/2 were significantly elevated in OE-DoMYB37 N. tabacum (Fig. 3C). The soluble phosphorus content in the above-ground part of the OE-DoMYB37 group was reduced by 0.72-fold, while the soluble phosphorus content in the below-ground part was reduced by 0.81-fold compared to the emptor vector (Fig. 3D). This indicates that overexpression of DoMYB37 reduces phosphate content, which promotes the expression of phosphate transporter proteins NtPT1/ NtPT2 and changes in root conformation.

Fig. 3
figure 3

Root length and related gene expression in OE-DoMYB37 transgenic N. tabacum and empty vector. A OE-DoMYB37 transgenic N. tabacum. B Root length of OE-DoMYB37 transgenic N. tabacum and emptor vector. C Expression levels of NtPT1 and NtPT2 in OE-DoMYB37 transgenic N. tabacum and emptor vector. D Soluble phosphorus content in OE-DoMYB37 transgenic N. tabacum and emptor vector. Significant differences between the Empty vector and OE-DoMYB37 seedlings grown under the same conditions (HP and LP) are indicated using asterisks (**p < 0.01; *p < 0.05; two-tailed Student’s t-test)

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In order to study the function of DoMYB37, we established a transient transformation system for the D. officinale protocorm-like bodies (Fig. 4A). We obtained the OE-DoMYB37 D. officinale protocorm-like bodies, LP promoted the expression level of DoMYB37 in the control and overexpression groups (Fig. 4B). Soluble phosphorus content in OE-DoMYB37 was reduced (Fig. 4C). The expression levels of the acid phosphatase genes (DoPAP17, DoPAP2) and DoPHT2 became higher in OE-DoMYB37 transgenic D. officinale protocorm-like bodies and were induced by LP (Fig. 4D). In addition, we found that genes related to glucomannan metabolism (DoGMP, DoPMM, and DoCSLA6) expressed at higher levels in overexpressing line than in the control group, and the expression levels of these genes were higher in the LP treatment compared to the HP (Fig. 4D). D. officinale protocorm-like bodies with transiently transformed DoMYB37 had lower phosphorus content than the control group. The above results showed that overexpression of DoMYB37 decreased the phosphorus content. Then the expression levels of the DoPAP17, DoPAP2 and DoPHT2 were increased, which was similar to the results of overexpression of DoSPX1.

Fig. 4
figure 4

Physiological indices in OE-DoMYB37 D. officinale protocorm-like bodies and control group. A D. officinale protocorm-like bodies, B Expression levels of DoMYB37, C Soluble phosphorus content, D Expression levels of PSI genes (DoPHT2, DoPAP) and related to gluconeogenesis (DoGMP, DoPMM, DoCSLA6) in OE-DoMYB37 D. officinale protocorm-like bodies and control group. Significant differences between the Empty vector and OE-DoMYB37 seedlings grown under the same conditions (HP and LP) are indicated using asterisks (**p < 0.01; *p < 0.05; two-tailed Student’s t-test)

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DoMYB37 binds to the DoCSLA6 promoter and regulates DoCSLA6 expression

DoCSLA6 may be involved in LP-induced mannose accumulation [10]. In addition, DoCSLA6 expression was significantly higher in D. officinale protocorm-like bodies overexpressing DoMYB37. Analysis of DoCSLA6 promoter sequence revealed a large number of MYB binding sites. Three fragments containing MBS elements (a binding site for the MYB transcription factor) in the DoCSLA6 promoter region (Fig. 5A). Yeast one-hybrid (Y1H) assays showed that DoMYB37 could bind the three MBS sites on the DoCSLA6 promoter. (Fig. 5B). The results of EMSA experiments showed that DoMYB37 can combine with the promoter regions of the DoCSLA6 (Fig. 5C). We transiently co-transformed DoSPX1 and DoMYB37 into protocorm-like bodies and found that DoCSLA6 expression was more higher in the DoSPX1 and DoMYB37 co-transformed group than MYB37 overexpressed alone (Fig. 5D). This suggests that DoSPX1 may work in concert with DoMYB37 to regulate the expression of DoCSLA6.

Fig. 5
figure 5

Interaction between DoMYB37 and proDoCSLA6, and determination of DoCSLA6 expression and polysaccharide content in transgenic protocorm-like bodies and control group. A Validation of yeast monohybrid crosses. B Yeast one-hybrid between DoMYB37 and proDoCSLA6. C EMSA between DoMYB37 and proDoCSLA6. D The expression of DoCSLA6 in transgenic protocorm-like bodies. Significant differences between the Empty vector, OE-DoMYB37 and OE-DoMYB37/DoSPX1 seedlings grown under the same conditions (HP and LP) are indicated using asterisks (**p < 0.01; *p < 0.05; two-tailed Student’s t-test)

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Discussion

Plants usually face LP enviroment, which limits their growth and development [20].The protein-containing SPX domain is vital for the regulation of Pi signaling and Pi homeostasis, the SPX proteins sense phosphorus deficiency signaling and transduction the signal [21]. Duan’s research shows that both AtSPX1 and AtSPX3 play positive roles in plant adaptation to phosphate starvation, and AtSPX3 may have a negative feedback regulatory role in AtSPX1 response to phosphate starvation [22]. We found that DoSPX1 has a functional structural domain that reduces phosphate content in plants. DoSPX1 can reduce phosphate accumulation in plants and promote the expression of PSI genes, thereby enhancing plant tolerance to LP environments.

MYB proteins play important roles in controlling metabolism regulation during the whole processes of plant growth and development. MYB TFs can be divided into four categories, namely, R1R2R3-MYB, R2R3-MYB, R1-MYB, and 4R-MYB [23]. R2R3-MYB S20 subfamily (MYB-S20) transcription factors have been reported to be central regulators of plant responses to the LP pathway. MYB-S20 can sense the phosphorus signal transmitted by SPX [24, 25]. AtMYB62 is an R2R3-type MYB transcription factor. It connects Pi homeostasis and GA signaling during Pi starvation [26]. DoMYB37 had the highest homology with AtMYB62 and the expression of DoMYB37 was significantly increased in LP environment. DoMYB37 may involed in phosphate signal in D. officinale.

In plants, SPX can interact with MYB, and then affects the transcriptional regulation of downstream phosphorus starvation-induced genes by MYB transcription factors. Previous studies indicated that rice SPX1 interacts with PHR2 (MYB-CC) in a Pi-dependent manner and acts as an inhibitor to repress PHR2 transcription activation under Pi-replete conditions [27]. We verified that DoSPX1 can interact with DoMYB37 and affect the content of plant phosphates. By overexpressing DoMYB37 in tobacco and D. officinale protocorm-like bodies, we found that the expression of PSI genes was significantly up-regulated. However, whether the interaction of DoSPX1 with DoMYB37 affects the transcriptional regulation of these genes by DoMYB37 needs to be further explored.

The MYB family can response to stress signal like temperature, light and diseases, and then play transcriptional function in plant metabolic pathways by binding to the MBS sites of critical enzyme gene promoters [28, 29]. Metabolic pathways that MYB transcription factors can regulate include anthocyanins in Arabidopsis thaliana [30], Sugar in rice [31], salvianolic acid in Salvia miltiorrhiza [28], paclitaxel biosynthesis in Taxus chinensis [32] and so on. Our previous study showed that DoCSLA6 expression was elevated in LP. By yeast one hybridization and EMSA analysis, we found DoCSLA6 is a target gene of DoMYB37. Expression of DoCSLA6 was significantly increased in overexpressing DoMYB37 D. officinale protocorm-like bodies. The expression level of the DoCSLA6 gene in the co-transformation of DoSPX1 and DoMYB37 was much higher than that in the DoMYB37 alone transformation. It suggests that DoSPX1 and DoMYB37 coregulated the expression of the DoCSLA6 gene. However, whether the increase in the expression of DoCSLA6 would affect the polysaccharides, mannose and glucose content of D. officinale needs to be further explored.

Conclusions

In D. officinale, both DoSPX1 and DoMYB37 respond to LP. Under LP environment, the early LP response factor DoSPX1 was interact with DoMYB37, DoMYB37 belong to S20 subfamily of MYB-R2R3 fimaly. DoMYB37 can bind the MBS site upstream of the DoCSLA6 promoter and promote the expression of DoCSLA6. In the presence of DoSPX1, the promotion effect of DoCSLA6 expression is more obvious. In summary, we established the signal transduction model (“DoSPX1-DoMYB37-DoCSLA6”) of D. officinale in response to LP. The DoCSLA6 can catalyzes the synthesis of glucomannan from GDP-glucose and GDP-mannose, we infered that the increase of DoCSLA6 expression may be the main reason for the increase of glucomannan,which needs to be further verified (Fig. 6).

Fig. 6
figure 6

DoSPX1 and DoMYB37 regulation of DoCSLA6 expression in low phosphorus

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Availability of data and materials

The raw data generated in this study are available in Mendeley Data (https://doi.org/10.17632/tddc3sws2k.1). All data supporting the findings of this study are available from the corresponding authors upon request.

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Acknowledgements

We thank Anhui Provincial Engineering Research Center for Development and Utilization of Local Characteristic Plant Resources for providing materials.

Funding

This work was supported by Natural Science Foundation of Hefei (2022036) and Nature Science Research Project of Anhui province (2108085MC80).

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Author notes

  1. ZhiYuan Feng and YaWen Li contributed equally to this work.

Authors and Affiliations

  1. School of Life Sciences, Anhui Agricultural University, Hefei, Anhui, 230036, People’s Republic of China

    ZhiYuan Feng, YaWen Li, SiXue Zhang, Jingjing Song, HaoXin Xiang, JunRu Huang, HongHong Fan & Lin Liu

  2. Integrated Experimental Station in Dabie Mountains, Anhui Agricultural University, Lu’an, China

    HongHong Fan

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  1. ZhiYuan Feng
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Contributions

LL, JJ and ZY led the writing of the manuscript; YW, HX performed the experiment and collected the data;JR, HH and LL analysed and discussed the data; SX checked the grammar of the manuscript. All authors contributed critically to the drafts.

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Correspondence to HongHong Fan or Lin Liu.

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Supplementary Information

Supplementary Material 1.

12870_2024_5512_MOESM2_ESM.xlsx

Supplementary Material 2: Supplementary Table 1. MS liquid medium formula. Supplementary Table 2. Name and sequence of the primers.

12870_2024_5512_MOESM3_ESM.docx

Supplementary Material 3: Supplementary Fig 1 Transformation of D. officinale protocorm-like bodies. Supplementary Fig 2. Subcellular localization of DoSPX1.Scale bars: 20μm.. Supplementary Fig 3. Expression of DoMYB37 at different phosphorus concentrations.

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Feng, Z., Li, Y., Zhang, S. et al. DoSPX1 and DoMYB37 regulate the expression of DoCSLA6 in Dendrobium officinale during phosphorus starvation. BMC Plant Biol 24, 803 (2024). https://doi.org/10.1186/s12870-024-05512-8

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Keywords

BMC Plant Biology

ISSN: 1471-2229

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