Scientia Agricultura Sinica ›› 2022, Vol. 55 ›› Issue (2): 365-377.doi: 10.3864/j.issn.0578-1752.2022.02.011

• HORTICULTURE • Previous Articles     Next Articles

Inhibiting Eeffect of Root-Cutting and Top-Pinching on Graft Healing of Tomato

CUI QingQing1,2(),MENG XianMin1,DUAN YunDan1,ZHUANG TuanJie1,DONG ChunJuan1,GAO LiHong2,SHANG QingMao1()   

  1. 1Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences/Key Laboratory of Biology and Genetic Improvement of Horticultural Crops, Ministry of Agriculture, Beijing 100081
    2College of horticulture, China Agricultural University/Beijing Key Laboratory of Growth and Developmental Regulation for Protected Vegetable Crops, Beijing 100193
  • Received:2021-03-17 Accepted:2021-08-10 Online:2022-01-16 Published:2022-01-26
  • Contact: QingMao SHANG E-mail:cuiqq_caas@163.com;shangqingmao@caas.cn

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Abstract:

【Objective】The root-cutting grafting seedlings and top-pinching double-stem grafting seedlings or multi-stem grafting seedlings have been widely used in tomato cultivation, which have significant effects on high yield. Understanding the mechanism of root-cutting and top-pinching on tomato graft healing could provide the theoretical basis for tomato grafting production. 【Method】Using ‘YingFen No. 8’ as scion and ‘ZhenAi No. 1’ as rootstock, the grafting healing process, survival rate and xylem connectivity of tomato after root-cutting and top-cutting were observed, and the contents of auxin and cytokinin as well as the expressions of healing related genes in the grafted part were measured. 【Result】Root-cutting and top-pinching inhibited the expression of xylem differentiation gene VND7 above the graft union, and significantly delayed xylem remodeling. The root-cutting treatment significantly decreased the contents of trans-Zeatin riboside (tZR) and trans-Zeatin (tZ) at 12 and 72 hours after grafting, as well as the contents of indole-3-acetic acid (IAA), methyl indole-3-acetate (ME-IAA) and indole-3-carboxaldehyde (ICAld) at 12 hours after grafting. The contents of IAA and ME-IAA at 12 hours after top-pinching grafting were significantly lower than those of the control and root-cutting treatment, and the expression level of the phloem differentiation related gene NEN4 at 3 and 12 hours after grafting was down-regulated. Root-cutting or top-pinching down-regulated the expression levels of cell division related genes Histone H4 and SlcycB1-4 above the graft union at 3-48 hours after grafting. Under root-cutting and top-pinching treatment, the contents of IAA and ME-IAA in grafting union were significantly lower than those under root-cutting treatment, and the contents of tZR and tZ and xylem connectivity were significantly lower than those under top-pinching treatment. The application of 10 mmol∙L -1 IAA and 10 mmol∙L-1 6-BA at the grafted union after top-pinching and root-cutting could significantly promote the xylem remodeling of grafted seedlings. 【Conclusion】In conclusion, root-cutting and top-pinching could reduce the accumulation of cytokinin and auxin, down-regulate the expression of genes related to healing, reduce xylem transduction capacity, and delay the process of tomato grafting healing.

Key words: tomato, graft, top-pinching, root-cutting, auxin, cytokinin

Table 1

Primer sequences for the Real-time PCR"

基因 Gene 登录号 Accession No. 引物序列（5′-3′） Primer sequence
SlIAA1 Solyc09g083280 SlIAA1-F: TGGATGGTGCCCCTTATCTA
SlIAA1-R: ACAAGAAGACATAAACATTTCCCAA
SlPIN1 Solyc03g118740 SlPIN1-F: GGCAATTGTACAGGCAGCTC
SlPIN1-R: CCAATGTAATCGGCAACGCA
SlHP1 Solyc01g080540 SlHP1-F: CAAGTGGAAATGGTGGGGAA
SlHP1-R: ACCATCCAAGAATCCCTCACG
VND7 Solyc06g065410 VND7-F: ACAACAATGAAGATGATCGTGGCG
VND7-R: CTGGGAAGCACTCAAGCAAATGG
NEN4 Solyc04g054720 NEN4-F: AACTGTCAAGGCACAACCAGC
NEN4-R: CCCAAGCTCTGTCTTGCATAAG
PXY Solyc03g093330 PXY-F: CTTATGGCTACATCGCACCTGAATA
PXY-R: TTCCATCGCCAAATCCTGGT
Histone H4 Solyc11g066160 Histone H4-F: GGGAGGCAAAGGATTAGGCA
Histone H4-R: GAACCCCACGAGTTTCCTCA
SlcycB1-4 Solyc01g009040 SlcycB1-4-F: AGAGGCGGGGAATAGTCGTC
SlcycB1-4-R: ACAGCAGGAGCAATCACAAGG
Actin41 Solyc04g011500 SlActin-F: CTTCCAGCAGATGTGGATTGC
SlActin-R: GCATCTCTGGTCCAGTAGGAAA

Fig. 1

Morphological observation of grafting seedlings after root-cutting and top-pinching CK: Control; P: Top-pinching; C: Root-cutting; PC: Scion top-pinching and rootstock root-cutting. The same as below"

Fig. 2

Effects of root-cutting and top-pinching on dry and fresh weight of tomato grafting seedlings Different lowercase letters indicate significant differences (P<0.05). The same as below"

Table 2

Effects of root-cutting and top-pinching on survival rate and wilting rate of tomato grafting seedlings"

处理
Treatment		 成活率
Survival rate (%)		 萎蔫率 Wilting rate (%)
6 d 7 d 8 d
CK 100±0.00a 2.78±0.03b 0.00±0.00b 0.00±0.00a
P 100±0.00a 8.33±0.00ab 2.78±0.03ab 0.00±0.00a
C 97.22±0.03a 11.11±0.06ab 5.56±0.03ab 2.78±0.03a
PC 94.44±0.03a 16.67±0.05a 11.11±0.03a 5.56±0.03a

Fig. 3

Healing process of tomato grafting after root-cutting and top-pinching Dashed box indicates the position of the graft union, with the scion (Sc) above the graft union and the rootstock (St) below the graft union. Pc, Co and Vb are short for parenchyma cell, cortex and vascular bundle, respectively"

Fig. 4

Improved effects of exogenous application of IAA and 6-BA on xylem remodeling in tomato grafting after root-cutting and top-pinching A: Xylem transport ability at 72 hours after grafting; dashed lines indicate the position of the cut, with the scion above the cut and the rootstock below the cut, and Vb is short for vascular bundle. B: Xylem transport ability at 120 hours after grafting. P0: Top-pinching; P1: Top-pinching + 10 mM IAA applied to the cut surfaces; P2: Top-pinching + 100 mmol∙L-1 IAA applied to the cut surfaces; P3: Top-pinching + foliar spray 10 mmol∙L-1 IAA; C0: Root-cutting; C1: Root-cutting + 1 mmol∙L-1 6-BA applied to the cut surfaces; C2: Root-cutting + 10 mmol∙L-1 6-BA applied to the cut surfaces; C3: Root-cutting + the hypocotyls were dipped in 0.1 mmol∙L-1 6-BA for 30 min"

Fig. 5

Changes of auxin content in graft union after root-cutting and top-pinching"

Fig. 6

Changes of cytokinin content in graft union after root-cutting and top-pinching"

Fig. 7

Expression of auxin and cytokinin response genes in graft union after root-cutting and top-pinching U: Above the graft union; D: Below the graft union. * indicate significant differences among various treatments and control (CK) at P<0.05. The same as below"

Fig. 8

Expression of genes related to grafting healing after root-cutting and top-pinching"

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