Tomato plant resistant to tomato yellow leaf curl virus, powdery mildew, and nematodes

A method for identifying and selecting a S. lycopersicum plant that is resistant to Tomato Yellow Leaf Curl Virus (TYLCV), powdery mildew (PM) and nematodes, wherein said method comprises:

• a) detecting
• the presence or absence of an allele of the OL4 gene conferring resistance to PM and nematodes, said allele of the OL4 gene being identified by SSR-OL4 marker detection,
• the presence or absence of an allele of the TY/gene conferring resistance to TYLCYV, said allele of TY/gene being identified by TO-0178067 and/or TY3-M3 marker detection, and
• the presence or absence of an allele of the Mi-1 gene conferring resistance to nematodes, said allele of Mi-1 gene being identified by Mi2.3 marker detection;
• and
• b) selecting as a plant resistant to TYLCV, PM and nematodes the S. lycopersicum plant in which the allele of the OL4 gene conferring resistance to PM and nematodes and the allele of the TY1 gene conferring resistance to TYLCV have been detected as present in coupling phase on chromosome 6 and the allele of the Mi-1 gene conferring resistance to nematodes has been detected as absent in said coupling phase on chromosome 6,
• wherein SSR-OL4 marker detection is performed by amplification, and detection of an amplification product of 382 bp, referred to as “allele of 382 bp” of the marker SSR-OL4, when using primers consisting of SEQ ID NO:
• 1 and SEQ ID NO:
• 2 indicates presence of the allele of the OL4 gene conferring resistance to PM and nematodes, and detection of an amplification product of 389 bp, referred to as “allele of 389 bp” of the marker SSR-OL4, when using primers consisting of SEQ ID NO:
• 1 and SEQ ID NO:
• 2 indicates absence of the allele of the OL4 gene conferring resistance to PM and nematodes;
• wherein TO-0178067 marker detection is performed by amplification, and detection of an adenine (A) rather than a guanine (G) at position 247 of SEQ ID NO:
• 3 in an amplification product comprising sequence SEQ ID NO:
• 3, or in a fragment thereof including a G/A polymorphism at position 247 of SEQ ID NO:
• 3, referred to as “allele A” of the TO-0178067 marker, indicates the presence of the allele of the TY1 gene conferring resistance to TYLCV, and detection of a guanine (G) rather than an adenine (A) at position 247 of SEQ ID NO:
• 3 in an amplification product comprising sequence SEQ ID NO:
• 3, or in a fragment thereof including the G/A polymorphism at position 247 of SEQ ID NO:
• 3, referred to as “allele G” of the TO-0178067 marker, indicates the absence of the allele of the TY1 gene conferring resistance to TYLCV;
• wherein TY3-M3 marker detection is performed by amplification, and detection of an amplification product of 220 bp, referred to as “allele of 220 bp” of the marker TY3-M3, when using primers consisting of SEQ ID NO:
• 7 and SEQ ID NO:
• 8 indicates presence of the allele of the TY1 gene conferring resistance to TYLCV, and detection of an amplification product of 160 bp, referred to as “allele of 160 bp” of the marker TY3-M3, when using primers consisting of SEQ ID NO:
• 7 and SEQ ID NO:
• 8, indicates absence of the allele of the TY1 gene conferring resistance to TYLCV;
• and
• wherein Mi2.3 marker detection is performed by amplification, and detection of a cytosine (C) rather than a guanine (G) at position 88 of SEQ ID NO:
• 9 in an amplification product comprising SEQ ID NO:
• 9, or in a fragment thereof including a C/G polymorphism at position 88 of SEQ ID NO:
• 9, referred to as “allele C” of the marker Mi2.3, indicates the absence of the allele of the Mi-1 gene conferring resistance to nematodes, and detection of a guanine (G) rather than a cytosine (C) at position 88 of SEQ ID NO:
• 9 in an amplification product comprising SEQ ID NO:
• 9, or in a fragment thereof including the C/G polymorphism at position 88 of SEQ ID NO:
• 9, referred to as “allele G” of the marker Mi2.3, indicates the presence of the allele of the Mi-1 gene conferring resistance to nematodes.

A method for obtaining S. lycopersicum plant resistant to Tomato Yellow Leaf Curl Virus (TYLCV), powdery mildew (PM) and nematodes which comprises crossing a S. lycopersicum that is resistant to Tomato Yellow Leaf Curl Virus (TYLCV), powdery mildew (PM) and nematodes, wherein said plant comprises (i) an allele of the OL4 gene conferring resistance to PM and nematodes and (ii) an allele of the TY/gene conferring resistance to TYLCV in coupling phase on chromosome 6, and wherein said plant, if it comprises an allele of the Mi-1 gene conferring resistance to nematodes, then said allele of the Mi-1 gene conferring resistance to nematodes is not in coupling phase with the allele of the OL4 gene conferring resistance to PM and nematodes and the allele of the TY1 gene conferring resistance to TYLCV on chromosome 6, with another S. lycopersicum plant,

• wherein:
• said allele of the OL4 gene conferring resistance to PM and nematodes is identified by SSR-OL4 marker detection,
• said allele of the TY1 gene conferring resistance to TYLCV is identified by TO-0178067 and/or TY3-M3 marker detection, and
• the presence or absence of the allele of the Mi-1 gene conferring resistance to nematodes is identified by Mi2.3 marker detection,
• wherein SSR-OL4 marker detection is performed by amplification, and detection of an amplification product of 382 bp, referred to as “allele of 382 bp” of the marker SSR-OL4, when using primers consisting of SEQ ID NO:
• 1 and SEQ ID NO:
• 2 indicates presence of the allele of the OL4 gene conferring resistance to PM and nematodes, and detection of an amplification product of 389 bp, referred to as “allele of 389 bp” of the marker SSR-OL4, when using primers consisting of SEQ ID NO:
• 1 and SEQ ID NO:
• 2 indicates absence of the allele of the OL4 gene conferring resistance to PM and nematodes;
• wherein TO-0178067 marker detection is performed by amplification, and detection of an adenine (A) rather than a guanine (G) at position 247 of SEQ ID NO:
• 3 in an amplification product comprising sequence SEQ ID NO:
• 3, or in a fragment thereof including a G/A polymorphism at position 247 of SEQ ID NO:
• 3, referred to as “allele A” of the TO-0178067 marker, indicates the presence of the allele of the TY1 gene conferring resistance to TYLCV, and detection of a guanine (G) rather than an adenine (A) at position 247 of SEQ ID NO:
• 3 in an amplification product comprising sequence SEQ ID NO:
• 3, or in a fragment thereof including the G/A polymorphism at position 247 of SEQ ID NO:
• 3, referred to as “allele G” of the TO-0178067 marker, indicates the absence of the allele of the TY1 gene conferring resistance to TYLCV;
• wherein TY3-M3 marker detection is performed by amplification, and detection of an amplification product of 220 bp, referred to as “allele of 220 bp” of the marker TY3-M3 when using primers consisting of SEQ ID NO:
• 7 and SEQ ID NO:
• 8 indicates presence of the allele of the TY1 gene conferring resistance to TYLCV, and detection of an amplification product of 160 bp, referred to as “allele of 160 bp” of the marker TY3-M3, when using primers consisting of SEQ ID NO:
• 7 and SEQ ID NO:
• 8, indicates absence of the allele of the TY/gene conferring resistance to TYLCV;
• and
• wherein Mi2.3 marker detection is performed by amplification, and detection of a cytosine (C) rather than a guanine (G) at position 88 of SEQ ID NO:
• 9 in an amplification product comprising SEQ ID NO:
• 9, or in a fragment thereof including a C/G polymorphism at position 88 of SEQ ID NO:
• 9, referred to as “allele C” of the marker Mi2.3, indicates the absence of the allele of the Mi-1 gene conferring resistance to nematodes, and detection of a guanine (G) rather than a cytosine (C) at position 88 of SEQ ID NO:
• 9 in an amplification product comprising SEQ ID NO:
• 9, or in a fragment thereof including the C/G polymorphism at position 88 of SEQ ID NO:
• 9, referred to as “allele G” of the marker Mi2.3, indicates the presence of the allele of the Mi-1 gene conferring resistance to nematodes.