Helicoverpa armigera single nucleopolyhedrovirus (HearSNPV) genotypes, method of producing same and use as a biological control agent

A method for identifying in a sample the presence of a genotype selected from HearSNPV-SP1 B (CNCM I-4806) and HearSNPV-LB8 (CNCM I-4807) of H. armigera single nucleopolyhedrovirus comprising the steps of:

• i) PCR amplification of DNA extracted from said sample using a pair of primers selected from the group consisting of those formed by:
• a) SEQ ID NO:
• 1 (F-hr1) and SEQ ID NO:2 (R-hr1), and
• b) SEQ ID NO:3 (F-hr5) and SEQ ID NO:4 (R-hr5);
• ii) analyzing the amplified fragment to determine its size or sequence;
• iii) digesting the amplified fragment using NdeI endonuclease;
• iv) analyzing the fragments generated after digestion with NdeI to determine the number of fragments and the size of each of them;
• v) concluding that one of the genotypes HearSNPV-SP1B (CNCM I-4806) or HearSNPV-LB6 (CNCM I-4807) is present if:
• a) the fragment amplified by the pair of SEQ ID NO:
• 1 and SEQ ID NO:
• 2 has:
• i) a length of 2177 (HearSNPV-SP1B) or 2117 (HearSNPV-LB6) nucleotides;
• ii) digestion of said fragment with NdeI endonuclease generates six fragments of 857, 508, 381, 306, 78 and 47 nucleotides (HearSNPV-SP1B) or five fragments of 1210, 475, 307, 78 and 47 nucleotides (HearSNPV-LB6);
• or
• iii) the sequence represented by SEQ ID NO:
• 5 (HearSNPV-SP1B) or SEQ ID NO:
• 6 (HearSNPV-LB6);
• or alternatively,
• b) the fragment amplified by the primer pair of SEQ ID NO:
• 3 and SEQ ID NO:
• 4 has:
• i) a length of 2326 (HearSNPV SP1B) or 2330 (HearSNPV-LB6) nucleotides;
• ii) digestion of the fragment with NdeI endonuclease generates four fragments of 1120, 917, 211 and 78 nucleotides (HearSNPV-SP1B) or three fragments of 1120, 998 and 212 nucleotides (HearSNPV-LB6);
• or
• iii) the sequence represented by SEQ ID NO:
• 7 (HearSNPV-SP1B) or SEQ ID NO:
• 8 (HearSNPV-LB6).

A composition comprising:

• i) HearSNPV deposited in the National Collection of Microorganism Cultures (CNCM) with the deposit numbers CNCM I-4806 (HearSNPV-SP1B) and CNCM I-4807 (HearSNPV-LB6), or
• ii) genotypes whose genome is represented by SEQ ID NO:
• 13 (HearSNPV-SP1B) and SEQ ID NO:
• 14 (HearSNPV-LB6).