Modulating climacteric intensity in melon through QTL stacking

Abstract

Fruit ripening is one of the main processes affecting fruit quality and shelf life. In melon there are both climacteric and non-climacteric genotypes, making it a suitable species to study fruit ripening. In the current study, in order to fine tune ripening, we have pyramided three climacteric QTLs in the non-climacteric genotype “Piel de Sapo”: ETHQB3.5, ETHQV6.3 and ETHQV8.1. The results showed that the three QTLs interact epistatically, affecting ethylene production and ripening-related traits such as aroma profile. Each individual QTL has a specific role in the ethylene production profile. ETHQB3.5 accelerates the ethylene peak, ETHQV6.3 advances the ethylene production and ETHQV8.1 enhances the effect of the other two QTLs. Regarding aroma, the three QTLs independently activated the production of esters changing the aroma profile of the fruits, with no significant effects in fruit firmness, soluble solid content and fruit size. Understanding the interaction and the effect of different ripening QTLs offers a powerful knowledge for candidate gene identification as well as for melon breeding programs, where fruit ripening is one of the main objectives.

References

[1] Seymour GB, Manning K, Eriksson EM et al. Genetic identification and genomic organization of factors affecting fruit texture. J Exp Bot. 2002; 53: 2065-71.

[2] Prasanna V, Prabha TN, Tharanathan RN . Fruit ripening phenomena-an overview. Crit Rev Food Sci Nutr. 2007; 47: 1-19.

[3] Giovannoni JJ . Fruit ripening mutants yield insights into ripening control. Curr Opin Plant Biol. 2007; 10: 283-9.

[4] Giovannoni JJ . Genetic regulation of fruit development and ripening. Plant Cell. 2004; 16: S170-80.

[5] Ezura H, Owino WO . Melon, an alternative model plant for elucidating fruit ripening. Plant Sci. 2008; 175: 121-9.

[6] Pech JC, Bouzayen M, Latché A . Climacteric fruit ripening: ethylene-dependent and independent regulation of ripening pathways in melon fruit. Plant Sci. 2008; 175: 114-20.

[7] Périn C, Hagen L, de Conto V et al. A reference map of Cucumis melo based on two recombinant inbred line populations. Theor Appl Genet. 2002; 104: 1017-34.

[8] Pereira L, Ruggieri V, Pérez S et al. QTL mapping of melon fruit quality traits using a high-density GBS-based genetic map. BMC Plant Biol. 2018;18:324.

[9] Eduardo I, Arús P, Monforte AJ . Development of a genomic library of near isogenic lines (NILs) in melon (Cucumis melo L.) from the exotic accession PI161375. Theor Appl Genet. 2005; 112: 139-48.

[10] Pereira L, Santo Domingo M, Argyris J et al. A novel introgression line collection to unravel the genetics of climacteric ripening and fruit quality in melon. Sci Rep. 2021; 11: 11364.

[11] Garcia-Mas J, Benjak A, Sanseverino W et al. The genome of melon (Cucumis melo L.). Proc Natl Acad Sci U S A. 2012; 109: 11872-7.

[12] Bernacchi D, Beck-Bunn T, Emmatty D et al. Advanced backcross QTL analysis of tomato. II. Evaluation of near-isogenic lines carrying single-donor introgressions for desirable wild QTL-alleles derived from Lycopersicon hirsutum and L. pimpinellifolium. Theor Appl Genet. 1998; 97: 170-80.

[13] Urrutia M, Rambla JL, Alexiou KG et al. Genetic analysis of the wild strawberry (Fragaria vesca) volatile composition. Plant Physiol Biochem. 2017; 121: 99-117.

[14] Serra O, Donoso JM, Picañol R et al. Marker-assisted introgression (MAI) of almond genes into the peach background: a fast method to mine and integrate novel variation from exotic sources in long intergeneration species. Tree Genet Genomes. 2016; 12: 96.

[15] Castro G, Perpiñá G, Monforte AJ et al. New melon introgression lines in a Piel de Sapo genetic background with desirable agronomical traits from dudaim melons. Euphytica. 2019; 215: 169.

[16] Perpiñá G, Cebolla-Cornejo J, Esteras C et al. ‘MAK-10’: a long shelf-life Charentais breeding line developed by introgression of a genomic region from Makuwa melon. HortScience. 2017; 52: 1633-8.

[17] Moreno E, Obando JM, Dos-Santos N et al. Candidate genes and QTLs for fruit ripening and softening in melon. Theor Appl Genet. 2008; 116: 589-602.

[18] Vegas J, Garcia-Mas J, Monforte AJ . Interaction between QTLs induces an advance in ethylene biosynthesis during melon fruit ripening. Theor Appl Genet. 2013; 126: 1531-44.

[19] Pereira L, Santo Domingo M, Ruggieri V et al. Genetic dissection of climacteric fruit ripening in a melon population segregating for ripening behavior. Hortic Res. 2020; 7: 187-18.

[20] Pereira L. Genetic dissection of fruit quality and ripening traits in melon. PhD thesis, Universitat Autònoma de Barcelona, Barcelona, 2018.

[21] Ríos P, Argyris J, Vegas J et al. ETHQV6.3 is involved in melon climacteric fruit ripening and is encoded by a NAC domain transcription factor. Plant J. 2017; 91: 671-83.

[22] Mayobre C, Pereira L, Eltahiri A et al. Genetic dissection of aroma biosynthesis in melon and its relationship with climacteric ripening. Food Chem. 2021; 353: 129484.

[23] Argyris JM, Pujol M, Martín-Hernández AM et al. Combined use of genetic and genomics resources to understand virus resistance and fruit quality traits in melon. Physiol Plant. 2015; 155: 4-11.

[24] Sandhu N, Dixit S, Swamy BPM et al. Marker assisted breeding to develop multiple stress tolerant varieties for flood and drought prone areas. Rice. 2019; 12: 8.

[25] Guan H, Hou X, Jiang Y et al. Feature of blast resistant near-isogenic lines using an elite maintainer line II-32B by marker-assisted selection. J Plant Pathol. 2019; 101: 491-501.

[26] Lavaud C, Baviere M, le Roy G et al. Single and multiple resistance QTL delay symptom appearance and slow down root colonization by Aphanomyces euteiches in pea near isogenic lines. BMC Plant Biol. 2016; 16: 166.

[27] Ortega MA, All JN, Boerma HR et al. Pyramids of QTLs enhance host-plant resistance and Bt-mediated resistance to leaf-chewing insects in soybean. Theor Appl Genet. 2016; 129: 703-15.

[28] Robbins MD, Casler MD, Staub JE . Pyramiding QTL for multiple lateral branching in cucumber using inbred backcross lines. Mol Breeding. 2008; 22: 131-9.

[29] Gur A, Zamir D . Mendelizing all components of a pyramid of three yield QTL in tomato. Front Plant Sci. 2015; 6: 1096.

[30] Zhang N, Xu BH, Bi YF et al. Development of a muskmelon cultivar with improved resistance to gummy stem blight and desired agronomic traits using gene pyramiding. Czech J Genet Plant Breed. 2017; 53: 23-9.

[31] Wang R, Lammers M, Tikunov Y et al. The rin, nor and Cnr spontaneous mutations inhibit tomato fruit ripening in additive and epistatic manners. Plant Sci. 2020; 294: 110436.

[32] Zhao G, Lian Q, Zhang Z et al. A comprehensive genome variation map of melon identifies multiple domestication events and loci influencing agronomic traits. Nat Genet. 2019; 51: 1607-15.

[33] Vrebalov J, Ruezinsky D, Padmanabhan V et al. A MADS-box gene necessary for fruit ripening at the tomato ripening-inhibitor (Rin) locus. Science. 2002; 296: 343-6.

[34] Manning K, TÖr M, Poole M et al. A naturally occurring epigenetic mutation in a gene encoding an SBP-box transcription factor inhibits tomato fruit ripening. Nat Genet. 2006; 38: 948-52.

[35] Saladié M, Cañizares J, Phillips MA et al. Comparative transcriptional profiling analysis of developing melon (Cucumis melo L.) fruit from climacteric and non-climacteric varieties . BMC Genomics. 2015; 16: 440.

[36] Oren E, Tzuri G, Dafna A et al. QTL mapping and genomic analyses of earliness and fruit ripening traits in a melon recombinant inbred lines population supported by de novo assembly of their parental genomes. Hortic Res. 2022; 9: uhab081.

[37] Obando-Ulloa JM, Moreno E, García-Mas J et al. Climacteric or non-climacteric behavior in melon fruit: 1. Aroma volatiles. Postharvest Biol Technol. 2008; 49: 27-37.

[38] Yahyaoui FEL, Wongs-Aree C, Latché A et al. Molecular and biochemical characteristics of a gene encoding an alcohol acyl-transferase involved in the generation of aroma volatile esters during melon ripening. Eur J Biochem. 2002; 269: 2359-66.

[39] Chatzopoulou F, Sanmartin M, Mellidou I et al. Silencing of ascorbate oxidase results in reduced growth, altered ascorbic acid levels and ripening pattern in melon fruit. Plant Physiol Biochem. 2020; 156: 291-303.

[40] Lu J, Hou J, Ouyang Y et al. A direct PCR-based SNP marker-assisted selection system (D-MAS) for different crops. Mol Breeding. 2020; 40: 9.

[41] Doyle J. DNA Protocols for Plants. In: Hewitt GM, Johnston AWB, Young JPW, eds. Molecular Techniques in Taxonomy. Springer Berlin Heidelberg: Berlin, Heidelberg, 1991, 283-93.

[42] Pereira L, Pujol M, Garcia-Mas J et al. Non-invasive quantification of ethylene in attached fruit headspace at 1 p.p.b. by gas chromatography-mass spectrometry. Plant J. 2017; 91: 172-83.

[43] R Core Team (2020). R: A Language and Environment for Statistical Computing . R Foundation for Statistical Computing, Vienna, Austria. https://www.gbif.org/tool/81287/r-a-language-and-environment-for-statistical-computing.