Growth Differences Among Grafted Seedlings with Two Rootstocks of Catalpa bungei and Comparative Analysis of Transcriptome

doi:10.13560/j.cnki.biotech.bull.1985.2022-1557

Abstract

Abstract:

Grafting is the main way of reproducing and expanding Catalpa bunge of superior cultivar. In order to investigate the growth differences of two ‘Suqiu No. 1’ superior cultivars, taking Catalpa ovata and Catalpa fargesii as rootstocks, the survival rate, new shoot length, new shoot thickness, scion diameter/stock diameter, leaf area of the two cultivars were studied for screening the better rootstock. Transcriptome sequencing was performed on the leaves and terminal buds of the two grafted seedlings by high-throughput sequencing platform. The differentially expressed genes were confirmed by quantitative real-time PCR. The results showed that the survival rate and the growth of the grafted seedlings with C. ovata as the rootstock were significantly higher than those with C. fargesii as the rootstock. Total 559 differentially expressed genes were screened by transcriptome sequencing, including total 192 up-regulated genes and total 367 down-regulated genes. GO enrichment analysis showed that total 559 differentially expressed genes were significantly enriched in metabolic process, cellular process, organelles, binding and catalytic activities; KEGG enrichment analysis showed that total 213 differentially expressed genes were significantly enriched to total 66 metabolic pathways in leaves, and total 137 differentially expressed genes were significantly enriched to total 45 metabolic pathways in terminal buds. The RT-qPCR trends of four differentially expressed genes（Unigene0040270, Unigene0006320, Unigene0036149 and Unigene0007805）were consistent with the transcriptome data. The results of this study showed that there were significant differences in the growth of the two grafted seedlings, among which the growth parameters of the grafted seedlings with C. fargesii as the rootstock were better. At the same time, the differences were preliminarily analyzed from the perspective of transcriptome, providing a certain reference value for the study of C. bungei grafting molecules.

Key words: grafting, Catalpa bungei, growth differences, transcriptome, differentially expressed genes

FU Yu, JIA Rui-rui, HE He, WANG Liang-gui, YANG Xiu-lian. Growth Differences Among Grafted Seedlings with Two Rootstocks of Catalpa bungei and Comparative Analysis of Transcriptome[J]. Biotechnology Bulletin, 2023, 39(8): 251-261.

Figures/Tables 13

References 37

[1]	Liu YM, Zhou L, Zhu YQ, et al. Anatomical features and its radial variations among different Catalpa bungei clones[J]. Forests, 2020, 11(8): 824. doi: 10.3390/f11080824 URL
[2]	Lu N, Zhang MM, Xiao Y, et al. Construction of a high-density genetic map and QTL mapping of leaf traits and plant growth in an interspecific F1 population of Catalpa bungei × Catalpa duclouxii Dode[J]. BMC Plant Biol, 2019, 19(1): 596. doi: 10.1186/s12870-019-2207-y
[3]	Wu JW, Li JY, Su Y, et al. A morphophysiological analysis of the effects of drought and shade on Catalpa bungei plantlets[J]. Acta Physiol Plant, 2017, 39(3): 80. doi: 10.1007/s11738-017-2380-2 URL
[4]	Xiao Y, Wang JH, Yun HL, et al. Genetic evaluation and combined selection for the simultaneous improvement of growth and wood properties in Catalpa bungei clones[J]. Forests, 2021, 12(7): 868. doi: 10.3390/f12070868 URL
[5]	王改萍, 王良桂, 王晓聪, 等. 楸树嫩枝扦插生根发育及根系特征分析[J]. 南京林业大学学报: 自然科学版, 2020, 44(6): 94-102.
	Wang GP, Wang LG, Wang XC, et al. Dynamic characteristics of cutting rooting of Catalpa bungei with tender branches[J]. J Nanjing For Univ Nat Sci Ed, 2020, 44(6): 94-102.
[6]	Chen W, Meng PP, Feng H, et al. Effects of arbuscular mycorrhizal fungi on growth and physiological performance of Catalpa bungei C.A.Mey. under drought stress[J]. Forests, 2020, 11(10): 1117. doi: 10.3390/f11101117 URL
[7]	Lin J, Wu LH, Jing L, et al. Effect of different plant growth regulators on callus induction in Catalpa bungei[J]. Afr J Agric Res, 2010, 5: 2699-2704.
[8]	Miao L, Li Q, Sun TS, et al. Sugars promote graft union development in the heterograft of cucumber onto pumpkin[J]. Hortic Res, 2021, 8(1): 146. doi: 10.1038/s41438-021-00580-5
[9]	Ren Y, Xu Q, Wang LW, et al. Involvement of metabolic, physiological and hormonal responses in the graft-compatible process of cucumber/pumpkin combinations was revealed through the integrative analysis of mRNA and miRNA expression[J]. Plant Physiol Biochem, 2018, 129: 368-380. doi: 10.1016/j.plaphy.2018.06.021 URL
[10]	Karaca F, Yetİşİr H, Solmaz İ, et al. Rootstock potential of Turkish Lagenaria siceraria germplasm for watermelon: plant growth, yield and quality[J]. Turkish J Agric For, 2012: 167-177.
[11]	Rasool A, Mansoor S, Bhat KM, et al. Mechanisms underlying graft union formation and rootstock scion interaction in horticultural plants[J]. Front Plant Sci, 2020, 11: 590847. doi: 10.3389/fpls.2020.590847 URL
[12]	Świerczyński S. The effect of rootstock activity for growth and root system soaking in Trichoderma atroviride on the graft success and continued growth of beech(Fagus sylvatica L.) plants[J]. Agronomy, 2022, 12(6): 1259. doi: 10.3390/agronomy12061259 URL
[13]	Liu WQ, Xiang CG, Li XJ, et al. Identification of long-distance transmissible mRNA between scion and rootstock in cucurbit seedling heterografts[J]. Int J Mol Sci, 2020, 21(15): 5253. doi: 10.3390/ijms21155253 URL
[14]	Kaseb MO, Umer MJ, Anees M, et al. Transcriptome profiling to dissect the role of genome duplication on graft compatibility mechanisms in watermelon[J]. Biology, 2022, 11(4): 575. doi: 10.3390/biology11040575 URL
[15]	Liu XY, Li J, Liu MM, et al. Transcriptome profiling to understand the effect of Citrus rootstocks on the growth of ‘shatangju’ mandarin[J]. PLoS One, 2017, 12(1): e0169897. doi: 10.1371/journal.pone.0169897 URL
[16]	Davoudi M, Song MF, Zhang MR, et al. Long-distance control of the scion by the rootstock under drought stress as revealed by transcriptome sequencing and mobile mRNA identification[J]. Hortic Res, 2022, 9: uhab033. doi: 10.1093/hr/uhab033 URL
[17]	申妍颖, 李雪涵, 李飞鸿, 等. 中间砧‘南通小方柿’嫁接柿转录组测序及分析[J]. 分子植物育种, 2019, 17(5): 1454-1466.
	Shen YY, Li XH, Li FH, et al. Transcriptome sequencing and analysis of grafted persimmon with ‘Nantong-Xiaofangshi’ as the interstock[J]. Mol Plant Breed, 2019, 17(5): 1454-1466.
[18]	Li GF, Ma JJ, Tan M, et al. Transcriptome analysis reveals the effects of sugar metabolism and auxin and cytokinin signaling pathways on root growth and development of grafted apple[J]. BMC Genom, 2016, 17(1): 150. doi: 10.1186/s12864-016-2484-x URL
[19]	Young MD, Wakefield MJ, Smyth GK, et al. Gene ontology analysis for RNA-seq: accounting for selection bias[J]. Genome Biol, 2010, 11(2): R14. doi: 10.1186/gb-2010-11-2-r14 URL
[20]	Mao XZ, Cai T, Olyarchuk JG, et al. Automated genome annotation and pathway identification using the KEGG Orthology(KO)as a controlled vocabulary[J]. Bioinformatics, 2005, 21(19): 3787-3793. doi: 10.1093/bioinformatics/bti430 URL
[21]	杨英英, 赵林姣, 杨桂娟, 等. ‘麦缘锦楸’叶色表型RT-qPCR内参基因筛选及验证[J]. 林业科学研究, 2022, 35(1): 123-131.
	Yang YY, Zhao LJ, Yang GJ, et al. Selection and validation of reference genes for leaf color phenotype in ‘maiyuanjinqiu’, a Catalpa fargesii variety, by RT-qPCR[J]. For Res, 2022, 35(1): 123-131.
[22]	He W, Xie R, Wang Y, et al. Comparative transcriptomic analysis on compatible/incompatible grafts in citrus[J]. Hortic Res, 2022, 9: uhab072. doi: 10.1093/hr/uhab072 URL
[23]	钟亚琴. 嫁接西瓜响应低钾胁迫的生理和分子机制研究[D]. 武汉: 华中农业大学, 2019.
	Zhong YQ. Physiological and molecular mechanism of grafted watermelon in response to low potassium stress[D]. Wuhan: Huazhong Agricultural University, 2019.
[24]	Thomas A, Brauer D, Sauer T, et al. Cultivar influences early rootstock and scion survival of grafted black walnut[J]. J Am Pomol Soc, 2008, 62(1): 3-12.
[25]	Reig G, Zarrouk O, Font i Forcada C, et al. Anatomical graft compatibility study between apricot cultivars and different plum based rootstocks[J]. Sci Hortic, 2018, 237: 67-73. doi: 10.1016/j.scienta.2018.03.035 URL
[26]	Huang Y, Zhao LQ, Kong QS, et al. Comprehensive mineral nutrition analysis of watermelon grafted onto two different rootstocks[J]. Hortic Plant J, 2016, 2(2): 105-113.
[27]	Bhuiyan MFA, Rahim MA, Alam MS. Study on the growth of plants produced by epicotyl(stone)grafting with different rootstock scion combinations in mango[J]. J Agrofor Environ, 2010, 3(2): 163-166.
[28]	Harris ZN, Awale M, Bhakta N, et al. Multi-dimensional leaf phenotypes reflect root system genotype in grafted grapevine over the growing season[J]. GigaScience, 2021, 10(12): giab087. doi: 10.1093/gigascience/giab087 URL
[29]	Yang YJ, Lu XM, Yan B, et al. Bottle gourd rootstock-grafting affects nitrogen metabolism in NaCl-stressed watermelon leaves and enhances short-term salt tolerance[J]. J Plant Physiol, 2013, 170(7): 653-661.
[30]	Somkuwar RG, Bahetwar A, Khan I, et al. Changes in growth, photosynthetic activities, biochemical parameters and amino acid profile of Thompson Seedless grapes(Vitis vinifera L.)[J]. J Environ Biol, 2014, 35(6): 1157-1163. pmid: 25522520
[31]	Proebsting WM, Maggard SP, Guo WW. The relationship of thiamine to the Alt locus of Pisum sativum L[J]. J Plant Physiol, 1990, 136(2): 231-235. doi: 10.1016/S0176-1617(11)81671-8 URL
[32]	Sayed SA, Gadallah MAA. Effects of shoot and root application of thiamin on salt-stressed sunflower plants[J]. Plant Growth Regul, 2002, 36(1): 71-80. doi: 10.1023/A:1014784831387 URL
[33]	Xu Q, Guo SR, Li L, et al. Proteomics analysis of compatibility and incompatibility in grafted cucumber seedlings[J]. Plant Physiol Biochem, 2016, 105: 21-28. doi: 10.1016/j.plaphy.2016.04.001 URL
[34]	Kim SH, Kim YH, Ahn YO, et al. Downregulation of the lycopene ϵ-cyclase gene increases carotenoid synthesis via the β-branch-specific pathway and enhances salt-stress tolerance in sweetpotato transgenic calli[J]. Physiol Plant, 2013, 147(4): 432-442. doi: 10.1111/ppl.2013.147.issue-4 URL
[35]	Komatsu A, Takanokura Y, Moriguchi T, et al. Differential expression of three sucrose-phosphate synthase isoforms during sucrose accumulation in citrus fruits(Citrus unshiu Marc.)[J]. Plant Science, 1999, 140(2): 169-178. doi: 10.1016/S0168-9452(98)00217-9 URL
[36]	Zhu YJ, Moore PH. Sucrose accumulation in the sugarcane stem is regulated by the difference between the activities of soluble acid invertase and sucrose phosphate synthase[J]. Plant Physiol, 1997, 115(2): 609-616. doi: 10.1104/pp.115.2.609 pmid: 12223829
[37]	Pauly M, Keegstra K. Plant cell wall polymers as precursors for biofuels[J]. Curr Opin Plant Biol, 2010, 13(3): 304-311. doi: 10.1016/j.pbi.2009.12.009 URL

嫁接组合 Grafting combination	取样部位 Sampling part	编号 Numbering
苏楸1号/梓树 ‘Suqiu No. 1’/ Catalpa ovata	叶片Leaf	ZS-L
苏楸1号/梓树 ‘Suqiu No. 1’/ Catalpa ovata	顶芽Bud	ZS-B
苏楸1号/滇楸 ‘Suqiu No. 1’/Catalpa fargesii	叶片Leaf	DS-L
苏楸1号/滇楸 ‘Suqiu No. 1’/Catalpa fargesii	顶芽Bud	DS-B

嫁接组合 Grafting combination	取样部位 Sampling part	编号 Numbering
苏楸1号/梓树 ‘Suqiu No. 1’/ Catalpa ovata	叶片Leaf	ZS-L
苏楸1号/梓树 ‘Suqiu No. 1’/ Catalpa ovata	顶芽Bud	ZS-B
苏楸1号/滇楸 ‘Suqiu No. 1’/Catalpa fargesii	叶片Leaf	DS-L
苏楸1号/滇楸 ‘Suqiu No. 1’/Catalpa fargesii	顶芽Bud	DS-B

引物名称Primer name	序列Sequence（5'-3'）
CfMADH-F	AGCTTCCATTCTTTGCCTCA
CfMADH-R	TCCGAACAAAAGCAATACCC
Unigene0036149-F	AGGAACAGGAGAATGTGAAGAGG
Unigene0036149-R	CCGTCGTCACTTCTCGCAG
Unigene0007805-F	CAAAAAATGCTGGAAAAGTACCC
Unigene0007805-R	TGGTAAAATCGTGGTTCTCAAGTG
Unigene0040270-F	CCGCTTCATCTTGGTTCAGTG
Unigene0040270-R	AAGAAGGAAGATTTGCTGTTGGA
Unigene0006320-F	AGAAACACGAAAGTGAATGCTAAAC
Unigene0006320-R	GCACTCGGATGAAACGAAAAC

引物名称Primer name	序列Sequence（5'-3'）
CfMADH-F	AGCTTCCATTCTTTGCCTCA
CfMADH-R	TCCGAACAAAAGCAATACCC
Unigene0036149-F	AGGAACAGGAGAATGTGAAGAGG
Unigene0036149-R	CCGTCGTCACTTCTCGCAG
Unigene0007805-F	CAAAAAATGCTGGAAAAGTACCC
Unigene0007805-R	TGGTAAAATCGTGGTTCTCAAGTG
Unigene0040270-F	CCGCTTCATCTTGGTTCAGTG
Unigene0040270-R	AAGAAGGAAGATTTGCTGTTGGA
Unigene0006320-F	AGAAACACGAAAGTGAATGCTAAAC
Unigene0006320-R	GCACTCGGATGAAACGAAAAC

嫁接组合 Grafting combination	成活率 Survival rate/%	新梢长度 New shoot length/cm	新梢粗度 New shoot thickness/cm	接穗粗度/砧木粗度 Scion diameter/Stock diameter	叶面积 Leaf area/cm²
ZS	60.95±0.064**	107.51±12.25**	2.47±0.65**	0.822±0.029**	338.64±6.11**
DS	32.05±0.038	83.43±5.06	2.04±1.57	0.721±0.087	247.61±6.59