Discovery of the interfamily grafting capacity of Petunia, a floricultural species

Abstract

In grafting, an agricultural technique for propagating flower species and fruit trees, two plants are combined to exploit their beneficial characteristics, such as rootstock disease tolerance and vigor. Grafting incompatibility has been observed, however, between distantly related plant combinations, which limits the availability of plant resources. A high grafting capacity has been found in Nicotiana, belonging to Solanaceae, but not in Ipomoea nil, a Convolvulaceae species. Here, we found that Petunia hybrida, another solanaceous species, has similar ability of interfamily grafting, which indicates that interfamily grafting capability in Solanaceae is not limited to the genus Nicotiana. RNA sequencing-based comparative time-series transcriptomic analyses of Nicotiana benthamiana, I. nil, and P. hybrida revealed that N. benthamiana and P. hybrida share a common gene expression pattern, with continued elevated expression of the β-1,4-glucanase subclade gene GH9B3 observed after interfamily grafting. During self-grafting, GH9B3 expression in each species was similarly elevated, thus suggesting that solanaceous plants have altered regulatory mechanisms for GH9B3 gene expression that allow tissue fusion even with other species. Finally, we tested the effect of the β-1,4-glucanase inhibitor D-glucono-1,5-lactone, using glucose as a control, on the interfamily grafting usability of P. hybrida with Arabidopsis rootstock. Strong inhibition of graft establishment was observed only with D-glucono-1,5-lactone, thus suggesting the important role of GH9B3 in P. hybrida grafting. The newly discovered grafting compatibility of Petunia with different families enhances the propagation techniques and the production of flower plants.

References

[1] Stehmann JO, Lorenz-Lemke A, Freitas L et al. The genus Petunia. In: Gerats T, Strommer J, eds. Petunia: Evolutionary, Developmental and Physiological Genetics. Springer: New York, NY, 2009, 1-28.

[2] Ando T . Distribution of Petunia axillaris (Solanaceae) and its new subspecies in Argentina and Bolivia. Acta Phytotax Geobot. 1996; 47: 19-30.

[3] Segatto ALA, Ramos-Fregonezi AM, Bonatto SL et al. Molecular insights into the purple-flowered ancestor of garden petunias. Am J of Bot. 2014; 101: 119-27.

[4] Gerats T, Vandenbussche M . A model system for comparative research: petunia. Trends Plant Sci. 2005; 10: 251-6.

[5] Horsch RB, Fry JE, Hoffmann NL et al. A simple and general method for transferring genes into plants. Science. 1985; 227: 1229-31.

[6] Strommer J, Peters J, Zethof J et al. AFLP maps of Petunia hybrida: building maps when markers cluster. Theor Appl Genet. 2002; 105: 1000-9.

[7] Bombarely A et al. Insight into the evolution of the Solanaceae from the parental genomes of Petunia hybrida . Nature Plants. 2016; 2: 16074.

[8] Napoli C . Highly branched phenotype of the petunia dad1-1 mutant is reversed by grafting. Plant Physiol. 1996; 111: 27-37.

[9] Mudge K, Janick J, Scofield S et al. Ch. 9. A History of Grafting. In: Janick J, ed. Horticultural Reviews 35.John Wiley & Sons, Inc., 2009, 437-93.

[10] Lee JM, Oda M . Ch. 2. Grafting of Herbaceous Vegetable and Ornamental Crops. In: Janick J, ed. Horticultural Reviews, 28.John Wiley & Sons, Inc., 2003, 61-124.

[11] Hartmann HT, Kester DE . Plant Propagation: Principles and Practices . 3rd ed. Englewood Cliffs, NJ: Prentice-Hall; 1975.

[12] Andrews PK, Marquez CS . Graft Incompatibility. In: Janick J, ed. Horticultural Reviews 15.John Wiley & Sons, Inc., 1993, 183-231.

[13] Melnyk CW . Plant grafting: insights into tissue regeneration. Regeneration. 2016; 4: 3-14.

[14] Notaguchi M, Kurotani KI, Sato Y et al. Cell-cell adhesion in plant grafting is facilitated by β-1,4-glucanases. Science. 2020; 369: 698-702.

[15] Yoshida S, Cui S, Ichihashi Y et al. The haustorium, a specialized invasive organ in parasitic plants. Annu Rev Plant Biol. 2016; 67: 643-67.

[16] Ishida JK, Wakatake T, Yoshida S et al. Local auxin biosynthesis mediated by a YUCCA flavin monooxygenase regulates haustorium development in the parasitic plant Phtheirospermum japonicum . Plant Cell. 2016; 28: 1795-814.

[17] Wakatake T, Ogaea S, Yoshida S et al. An auxin transport network underlies xylem bridge formation between the hemi-parasitic plant Phtheirospermum japonicum and host Arabidopsis . Development. 2020; 147: dev187781.

[18] Kurotani K, Wakatake T, Ichihashi Y et al. Host-parasite tissue adhesion by a secreted type of β-1,4-glucanase in the parasitic plant Phtheirospermum japonicum . Commun Bio. 2020; 3: 407.

[19] Okayasu K, Aoki K, Kurotani K et al. Tissue adhesion between distant plant species in parasitism and grafting. Commun & Integ Biol. 2021; 14: 21-3.

[20] Matsuoka K, Yanagi R, Yumoto E et al. RAP2.6L and jasmonic acid-responsive genes are expressed upon Arabidopsis hypocotyl grafting but are not needed for cell proliferation related to healing. Plant Mol Biol. 2018; 96: 531-42.

[21] Melnyk CW, Gabel A, Hardcastle TJ et al. Transcriptome dynamics at Arabidopsis graft junctions reveal an intertissue recognition mechanism that activates vascular regeneration. Proc Natl Acad Sci U S A. 2018; 115: E2447-56.

[22] Asahina M, Azuma K, Pitaksaringkarn W et al. Spatially selective hormonal control of RAP2.6L and ANAC071 transcription factors involved in tissue Reunion in Arabidopsis. Proc Natl Acad Sci U S A. 2011; 108: 16128-32.

[23] Matsuoka K, Sugawara E, Aoki R et al. Differential cellular control by cotyledon-derived Phytohormones involved in graft Reunion of Arabidopsis hypocotyls. Plant Cell Physiol. 2016; 57: 2620-31.

[24] Deshpande MV, Eriksson K, Pettersson LG . An assay for selective determination of exo-1,4,-β-glucanases in a mixture of cellulolytic enzymes. Anal Biochem. 1984; 138: 481-7.

[25] Vandenbussche M, Chambrier P, Bento SR et al. Petunia, your next supermodel? Frontier Plant Sci . 2016; 7: 72.

[26] Conn CE, Bythell-Douglas R, Neumann D et al. Plant evolution. Convergent evolution of strigolactone perception enabled host detection in parasitic plants. Science. 2015; 349: 540-3.

[27] Nelissen H, Clarke JH, De Block M et al. DRL1, a homolog of the yeast TOT4/KTI12 protein, has a function in meristem activity and organ growth in plants. Plant Cell. 2003; 15: 639-54.

[28] Nelissen H, Fleury D, Bruno L et al. The elongate mutants identify a functional Elongator complex in plants with a role in cell proliferation during organ growth. Proc Natl Acad Sci. 2005; 102: 7754-9.

[29] Iwase A, Mitsuda N, Koyama T et al. The AP2/ERF transcription factor WIND1 controls cell differentiation in Arabidopsis. Curr Biol. 2011; 21: 508-14.

[30] Hoang N, Choe G, Zheng Y et al. Identification of conserved gene-regulatory networks that integrate environmental sensing and growth in the root cambium. Curr Biol. 2020; 30: 2887-900.

[31] Kohorn BD, Kobayashi M, Johansen S et al. An Arabidopsis cell wall-associated kinase required for invertase activity and cell growth. Plant J. 2006; 46: 307-16.

[32] Toruño TY, Shen M, Coaker G et al. Regulated disorder: posttranslational modifications control the RIN4 plant immune signaling hub. Mol Plant-Microbe Interact. 2019; 32: 56-64.

[33] Schmidt DD, Voelckel C, Hartl M et al. Specificity in ecological interactions. Attack from the same lepidopteran herbivore results in species-specific transcriptional responses in two Solanaceous host plants. Plant Physiol. 2005; 138: 1763-73.

[34] Hernandez I, Alegre L, Van Breusegem F et al. How relevant are flavonoids as antioxidants in plants? Trends in Plant Sci. 2009; 14: 125-32.

[35] Gebler J, Gilkes NR, Claeyssens M et al. Stereoselective hydrolysis catalyzed by related β-1,4-glucanases and β-1,4-xylanases. J Biol Chem. 1992; 25: 12559-61.

[36] Urbanowicz BR, Bennett AB, del Campillo E et al. Structural organization and a standardized nomenclature for plant endo-1,4-beta-glucanases (cellulases) of glycosyl hydrolase family 9. Plant Physiol. 2007; 144: 1693-6.

[37] Gilbert HJ . The biochemistry and structural biology of plant cell wall deconstruction. Plant Physiol. 2010; 153: 444-55.

[38] Libertini E, Li Y, McQueen-Mason SJ . Phylogenetic analysis of the plant endo-beta-1,4-glucanase gene family. J Mol Evo. 2004; 58: 506-15.

[39] del Campillo E, Abdel-Aziz A, Crawford D et al. Root cap specific expression of an endo-beta-1,4-D-glucanase (cellulase): a new marker to study root development in Arabidopsis. Plant Mol Biol. 2004; 56: 309-23.

[40] Yoshida S, Kim S, Wafula EK et al. Genome sequence of Striga asiatica provides insight into the evolution of plant parasitism. Curr Biol. 2019; 29: 3041-3052.e3044.

[41] Bolger AM, Lohse M, Usadel B . Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics. 2014; 30: 2114-20.

[42] Kim D, Paggi JM, Park C et al. Graph-based genome alignment and genotyping with HISAT2 and HISAT-genotype. Nat Biotechnol. 2019; 37: 907-15.

[43] Li H, Handsaker B, Wysoker A et al. The sequence alignment/map (SAM) format and SAMtools. Bioinformatics. 2009; 25: 2078-9.

[44] Trapnell C, Roberts A, Goff L et al. Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and cufflinks. Nat Protoc. 2012; 7: 562-78.

[45] Bombarely A, Rosli HG, Vrebalov J et al. A draft genome sequence of Nicotiana benthamiana to enhance molecular plant-microbe biology research. Mol Plant-Microbe Interact. 2012; 25: 1523-30.

[46] Huang DW, Sherman BT, Lempicki RA . Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nature Protoc. 2009; 4: 44-57.

[47] Huang DW, Sherman BT, Lempicki RA . Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists. Nucleic Acids Res. 2009; 37: 1-13.