Novel type of long shelf-life melon plants

Abstract

The present invention relates to novel melon plants producing fruits displaying a combination of a long shelf-life trait as well as a maturity indicator trait. The present invention also relates to seeds and parts of said plants, for example fruits. The present invention further relates to methods of making and using such seeds and plants. The present invention also relates to novel genetic sequences associated with a rind-turning phenotype at maturity, which, when combined with a long shelf-life allele, significantly alters the characteristics of the maturing melon fruit, functions as a reliable maturity indicator while retaining suitable marketable features and results in a novel melon plant type.

1 . A cultivated melon plant, preferably a cultivated Cantaloupe melon plant, more preferably a  Cucumis melo  var.  reticulatus  plant or a  Cucumis melo  var.  cantalupensis  plant comprising in its genome:
 a) at least one copy of an LSL10 allele, and;   b) at least one copy of an introgressed sequence from  C. melo  var.  dudaim  associated with the rind-turning phenotype, located on chromosome 6 and comprising at least one of the following SNP markers:
 i) an A genotype in the heterozygous or homozygous state for SNP marker 1 at a position corresponding to position 61 in SEQ ID NO: 1; 
 ii) a G genotype in the heterozygous or homozygous state for SNP marker 2 at a position corresponding to position 55 in SEQ ID NO: 6; 
 iii) a G genotype in the heterozygous or homozygous state for SNP marker 3 at a position corresponding to position 65 in SEQ ID NO: 11; 
 iv) a G genotype in the heterozygous or homozygous state for SNP marker 4 at a position corresponding to position 49 in SEQ ID NO: 16; 
 v) a C genotype in the heterozygous or homozygous state for SNP marker 5 at a position corresponding to position 53 in SEQ ID NO: 21; 
 vi) a G genotype in the heterozygous or homozygous state for SNP marker 6 at a position corresponding to position 83 in SEQ ID NO: 26; 
 vii) a G genotype in the heterozygous or homozygous state for SNP marker 7 at a position corresponding to position 103 in SEQ ID NO: 31; 
 viii) an A genotype in the heterozygous or homozygous state for SNP marker 8 at a position corresponding to position 115 in SEQ ID NO: 36; 
 ix) an A genotype in the heterozygous or homozygous state for SNP marker 9 at a position corresponding to position 87 in SEQ ID NO: 41; and/or 
 x) a C genotype in the heterozygous or homozygous state for SNP marker 10 at a position corresponding to position 196 in SEQ ID NO: 46; 
   wherein said plant produces melon fruit exhibiting a long shelf-life phenotype, and wherein said melon fruit further exhibits a rind-turning phenotype when reaching full maturity.   
     
     
         2 . The plant according to  claim 1 , wherein said rind-turning phenotype is characterized by a rind colour turn from green when immature into yellow when reaching full maturity. 
     
     
         3 . The plant according to  claim 2 , wherein said melon fruit rind yellow colour at full maturity ranges from 15 to 20 (A-D) when measured using the colour patch of the Royal Horticultural Society Colour Chart. 
     
     
         4 . The plant according to  claim 3 , wherein said melon fruit rind yellow colour at full maturity is 19A or 19B when measured using the colour patch of the Royal Horticultural Society Colour Chart. 
     
     
         5 . The plant according to  claim 1 , wherein said immature melon fruit rind green colour ranges from 135 to 143 (A-D) when measured using the colour patch of the Royal Horticultural Society Colour Chart. 
     
     
         6 . The plant according to  claim 1 , wherein said immature melon fruit rind green colour is 138C, 138D or 139A when measured using the colour patch of the Royal Horticultural Society Colour Chart. 
     
     
         7 . The plant according to  claim 1 , wherein the rind-turning phenotype is evaluated as disclosed in Example 2A. 
     
     
         8 . The plant according to  claim 1 , wherein:
 i) the A genotype for SNP marker 1 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 2 and reverse primer of SEQ ID NO: 5, and probe of SEQ ID NO: 3;   ii) the G genotype for SNP marker 2 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 7 and reverse primer of SEQ ID NO: 10, and probe of SEQ ID NO: 8;   iii) the G genotype for SNP marker 3 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 12 and reverse primer of SEQ ID NO: 15, and probe of SEQ ID NO: 13;   iv) the G genotype for SNP marker 4 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 17 and reverse primer of SEQ ID NO: 20, and probe of SEQ ID NO: 18;   v) the C genotype for SNP marker 5 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 22 and reverse primer of SEQ ID NO: 25, and probe of SEQ ID NO: 23;   vi) the G genotype for SNP marker 6 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 27 and reverse primer of SEQ ID NO: 30, and probe of SEQ ID NO: 28;   vii) the G genotype for SNP marker 7 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 32 and reverse primer of SEQ ID NO: 35, and probe of SEQ ID NO: 33;   viii) the A genotype for SNP marker 8 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 37 and reverse primer of SEQ ID NO: 40, and probe of SEQ ID NO: 38;   ix) the A genotype for SNP marker 9 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 42 and reverse primer of SEQ ID NO: 45, and probe of SEQ ID NO: 43; and/or   x) the C genotype for SNP marker 10 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 47 and reverse primer of SEQ ID NO: 50, and probe of SEQ ID NO: 48.   
     
     
         9 . The plant according to  claim 1 , wherein said introgressed sequence comprises at least one of SEQ ID NO: 1, SEQ ID NO: 6, SEQ ID NO: 11, SEQ ID NO: 16, SEQ ID NO: 21, SEQ ID NO: 26, SEQ ID NO: 31, SEQ ID NO: 36, SEQ ID NO: 41 and/or SEQ ID NO: 46, or a sequence that is at least 80% identical to one or more of said sequences. 
     
     
         10 . The plant according to  claim 1 , wherein said LSL10 allele comprises the following SNP marker:
 a) a G genotype in the heterozygous or homozygous state for SNP marker 11 at a position corresponding to position 107 in SEQ ID NO: 51.   
     
     
         11 . The plant according to  claim 10 , wherein
 a) the G genotype for SNP marker 11 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 52 and reverse primer of SEQ ID NO: 55, and probe of SEQ ID NO: 53.   
     
     
         12 . The plant according to  claim 10 , wherein said LSL10 allele comprises SEQ ID NO: 51, or a sequence that is at least 80% identical to said sequence. 
     
     
         13 . The plant according to  claim 1 , wherein said plant is homozygous for said LSL10 allele and/or for said introgressed sequence located on chromosome 6. 
     
     
         14 . The plant of  claim 1 , wherein said LSL10 allele and said introgressed sequence are comprised in melon line 19MNA106815, representative seed of which is deposited under ATCC Accession No. PTA-126875, or a progeny or an ancestor thereof. 
     
     
         15 . The plant of  claim 1 , wherein said plant is obtained by crossing melon line 19MNA106815, representative seed of which is deposited under ATCC Accession No. PTA-126875, or a progeny or an ancestor thereof, with a melon plant that does not contain said introgressed sequence. 
     
     
         16 . The plant of  claim 1  wherein said plant is an inbred, a dihaploid or a hybrid plant. 
     
     
         17 . A plant of melon line 19MNA106815, representative seed of which is deposited under ATCC Accession No. PTA-126875. 
     
     
         18 . A plant part of a plant according to  claim 1 . 
     
     
         19 . A seed that produces a plant or a plant part according to  claim 1 . 
     
     
         20 . A method for producing a cultivated melon plant, preferably a cultivated Cantaloupe melon plant, more preferably a  Cucumis melo  var.  reticulatus  plant or a  Cucumis melo  var.  cantalupensis  plant, wherein said plant produces melon fruits exhibiting a long shelf-life phenotype as well as a rind-turning phenotype when reaching full maturity, wherein the method comprising the steps of
 a) crossing a plant according to  claim 1  with a cultivated melon plant lacking said LSL10 allele and said introgressed sequence;   b) selecting a progeny plant comprising at least one copy of an LSL10 allele and at least one copy of an introgressed sequence from  C. melo  var.  dudaim  located on chromosome 6, said selecting step comprising detecting at least one of the following SNP markers:
 i) an A genotype in the heterozygous or homozygous state for SNP marker 1 at a position corresponding to position 61 in SEQ ID NO: 1; 
 ii) a G genotype in the heterozygous or homozygous state for SNP marker 2 at a position corresponding to position 55 in SEQ ID NO: 6; 
 iii) a G genotype in the heterozygous or homozygous state for SNP marker 3 at a position corresponding to position 65 in SEQ ID NO: 11; 
 iv) a G genotype in the heterozygous or homozygous state for SNP marker 4 at a position corresponding to position 49 in SEQ ID NO: 16; 
 v) a C genotype in the heterozygous or homozygous state for SNP marker 5 at a position corresponding to position 53 in SEQ ID NO: 21; 
 vi) a G genotype in the heterozygous or homozygous state for SNP marker 6 at a position corresponding to position 83 in SEQ ID NO: 26; 
 vii) a G genotype in the heterozygous or homozygous state for SNP marker 7 at a position corresponding to position 103 in SEQ ID NO: 31; 
 viii) an A genotype in the heterozygous or homozygous state for SNP marker 8 at a position corresponding to position 115 in SEQ ID NO: 36; 
 ix) an A genotype in the heterozygous or homozygous state for SNP marker 9 at a position corresponding to position 87 in SEQ ID NO: 41; and/or 
 x) a C genotype in the heterozygous or homozygous state for SNP marker 10 at a position corresponding to position 196 in SEQ ID NO: 46; 
   thereby producing a plant producing fruits with a long shelf-life phenotype as well as a rind-turning phenotype when reaching full maturity.   
     
     
         21 . The method according to  claim 20 , wherein the method further comprises:
 c) selfing the selected progeny or crossing the selected progeny with another melon plant to produce further progeny.   
     
     
         22 . The method according to  claim 21 , wherein further progeny is selected and selfed/crossed for 2 to 10 more generations. 
     
     
         23 . The method according to  claim 20  wherein the plant of step a) is melon line 19MNA106815, representative seed of which is deposited under ATCC Accession No. PTA-126875, or a progeny or an ancestor thereof. 
     
     
         24 . A method for producing a F1 melon plant exhibiting rind-turning phenotype, the method comprising crossing an inbred melon plant, which is a plant according to  claim 1 , with a different inbred melon plant to produce F1 hybrid progeny. 
     
     
         25 . A method for identifying a cultivated melon plant, preferably a cultivated Cantaloupe melon plant, more preferably a  Cucumis melo  var.  reticulatus  plant or a  Cucumis melo  var.  cantalupensis  plant, wherein said plant produces melon fruits exhibiting a long shelf life phenotype as well as a rind-turning phenotype when reaching full maturity, wherein said plant comprises at least one copy of an LSL10 allele and at least one copy of an introgressed sequence from  C. melo  var.  dudaim  located on chromosome 6, wherein said method comprising the step of detecting at least one of the following SNP markers:
 a) an A genotype in the heterozygous or homozygous state for SNP marker 1 at a position corresponding to position 61 in SEQ ID NO: 1;   b) a G genotype in the heterozygous or homozygous state for SNP marker 2 at a position corresponding to position 55 in SEQ ID NO: 6;   c) a G genotype in the heterozygous or homozygous state for SNP marker 3 at a position corresponding to position 65 in SEQ ID NO: 11;   d) a G genotype in the heterozygous or homozygous state for SNP marker 4 at a position corresponding to position 49 in SEQ ID NO: 16;   e) a C genotype in the heterozygous or homozygous state for SNP marker 5 at a position corresponding to position 53 in SEQ ID NO: 21;   f) a G genotype in the heterozygous or homozygous state for SNP marker 6 at a position corresponding to position 83 in SEQ ID NO: 26;   g) a G genotype in the heterozygous or homozygous state for SNP marker 7 at a position corresponding to position 103 in SEQ ID NO: 31;   h) an A genotype in the heterozygous or homozygous state for SNP marker 8 at a position corresponding to position 115 in SEQ ID NO: 36;   i) an A genotype in the heterozygous or homozygous state for SNP marker 9 at a position corresponding to position 87 in SEQ ID NO: 41; and/or   j) a C genotype in the heterozygous or homozygous state for SNP marker 10 at a position corresponding to position 196 in SEQ ID NO: 46;   thereby identifying a melon plant producing fruits with a long shelf-life phenotype as well as a rind-turning phenotype when reaching full maturity.   
     
     
         26 . The method according to  claim 25 , wherein the method further comprises the step of detecting the following SNP marker
 a) a G genotype in the heterozygous or homozygous state for SNP marker 11 at a position corresponding to position 107 in SEQ ID NO: 51.   
     
     
         27 . The method according to  claim 25 , wherein said method further comprises selecting a melon plant comprising said one or more SNP markers, and crossing the selected melon plant with a second melon plant to produce progeny melon plants that comprise at least one of said SNP markers and produces fruits with a long shelf-life phenotype as well as a rind-turning phenotype when reaching full maturity. 
     
     
         28 . A method of producing melon seed, the method comprising growing a melon plant from the seed of  claim 19  and allowing the plant to produce further melon seed. 
     
     
         29 . A method for assessing the genotype of a cultivated melon plant, preferably a cultivated Cantaloupe melon plant, more preferably a  Cucumis melo  var.  reticulatus  plant or a  Cucumis melo  var.  cantalupensis  plant, wherein said plant produces melon fruits exhibiting a long shelf-life phenotype as well as a rind-turning phenotype when reaching full maturity, said method comprising the steps of:
 a) providing a sample from said plant, and,   b) detecting in said sample a QTL locus located on chromosome 6 and associated with said long shelf-life phenotype as well as a rind-turning phenotype, said QTL locus being flanked by SNP markers 1 and 10, and at least one of the following SNP markers:
 i) an A genotype in the heterozygous or homozygous state for SNP marker 1 at a position corresponding to position 61 in SEQ ID NO: 1; 
 ii) a G genotype in the heterozygous or homozygous state for SNP marker 2 at a position corresponding to position 55 in SEQ ID NO: 6; 
 iii) a G genotype in the heterozygous or homozygous state for SNP marker 3 at a position corresponding to position 65 in SEQ ID NO: 11; 
 iv) a G genotype in the heterozygous or homozygous state for SNP marker 4 at a position corresponding to position 49 in SEQ ID NO: 16; 
 v) a C genotype in the heterozygous or homozygous state for SNP marker 5 at a position corresponding to position 53 in SEQ ID NO: 21; 
 vi) a G genotype in the heterozygous or homozygous state for SNP marker 6 at a position corresponding to position 83 in SEQ ID NO: 26; 
 vii) a G genotype in the heterozygous or homozygous state for SNP marker 7 at a position corresponding to position 103 in SEQ ID NO: 31; 
 viii) an A genotype in the heterozygous or homozygous state for SNP marker 8 at a position corresponding to position 115 in SEQ ID NO: 36; 
 ix) an A genotype in the heterozygous or homozygous state for SNP marker 9 at a position corresponding to position 87 in SEQ ID NO: 41; and/or 
 x) a C genotype in the heterozygous or homozygous state for SNP marker 10 at a position corresponding to position 196 in SEQ ID NO: 46; and/or 
 xi) any other DNA marker associated with said QTL locus flanked by SNP markers 1 and 10. 
   
     
     
         30 . A method of identifying in a cultivated melon plant, preferably a cultivated Cantaloupe melon plant, more preferably a  Cucumis melo  var.  reticulatus  plant or a  Cucumis melo  var.  cantalupensis  plant, an introgressed sequence associated with a rind-turning phenotype when reaching full maturity, said method comprising the step of detecting in said plant an allele of at least one DNA marker that is genetically linked to a QTL locus associated with said rind-turning phenotype, wherein said allele maps within 10 cM, preferably within 5 cM of said QTL locus located on chromosome 6 in a genomic region flanked by SNP markers 1 and 10. 
     
     
         31 . The method according to  claim 30 , wherein said QTL locus can be identified by at least one of the following SNP markers
 a) an A genotype in the heterozygous or homozygous state for SNP marker 1 at a position corresponding to position 61 in SEQ ID NO: 1;   b) a G genotype in the heterozygous or homozygous state for SNP marker 2 at a position corresponding to position 55 in SEQ ID NO: 6;   c) a G genotype in the heterozygous or homozygous state for SNP marker 3 at a position corresponding to position 65 in SEQ ID NO: 11;   d) a G genotype in the heterozygous or homozygous state for SNP marker 4 at a position corresponding to position 49 in SEQ ID NO: 16;   e) a C genotype in the heterozygous or homozygous state for SNP marker 5 at a position corresponding to position 53 in SEQ ID NO: 21;   f) a G genotype in the heterozygous or homozygous state for SNP marker 6 at a position corresponding to position 83 in SEQ ID NO: 26;   g) a G genotype in the heterozygous or homozygous state for SNP marker 7 at a position corresponding to position 103 in SEQ ID NO: 31;   h) an A genotype in the heterozygous or homozygous state for SNP marker 8 at a position corresponding to position 115 in SEQ ID NO: 36;   i) an A genotype in the heterozygous or homozygous state for SNP marker 9 at a position corresponding to position 87 in SEQ ID NO: 41; and/or   j) a C genotype in the heterozygous or homozygous state for SNP marker 10 at a position corresponding to position 196 in SEQ ID NO: 46.   
     
     
         32 . The method according to  claim 31  further comprising the step of selecting a cultivated melon plant, preferably a cultivated Cantaloupe melon plant, more preferably a  Cucumis melo  var.  reticulatus  plant or a  Cucumis melo  var.  cantalupensis  plant comprising said introgressed sequence. 
     
     
         33 . A method of identifying a cultivated melon plant, preferably a cultivated Cantaloupe melon plant, more preferably a  Cucumis melo  var.  reticulatus  plant or a  Cucumis melo  var.  cantalupensis  plant, exhibiting a long shelf-life phenotype as well as a rind-turning phenotype by identifying a QTL associated with said rind-turning phenotype, the method comprising the steps of:
 a) detecting at least one DNA marker from a melon plant, which DNA marker is linked to a chromosomal interval associated with rind-turning phenotype, wherein said chromosomal interval is flanked on each side by SNP markers having at least 80% sequence identity to SEQ ID NOs: 1 and 46; and   b) identifying said melon plant comprising said at least one DNA marker.   
     
     
         34 . A method of identifying a melon source of rind-turning trait on chromosome 6, comprising:
 a) providing a melon material or a plurality of melon materials,   b) screening said melon material or plurality of melon materials using a kit which detects at least one of the SNP markers listed in Table 4, and,   c) identifying a wild melon accession comprising said at least one SNP marker selected in the list of Table 4.   
     
     
         35 . Use of any of SEQ ID NOs 1-50 for screening a population of melon plants for the presence of a QTL locus located on chromosome 6 and associated with a rind-turning phenotype.