Studium rezistence bramboru vůči háďátku bramborovému a plísni bramborové na úrovni DNA markerů

Study of potato resistance against the potato cyst nematodes and late blight by means of DNA markers

Studium rezistence bramboru vůči háďátku bramborovému a plísni bramborové na úrovni DNA markerů

Petr SEDLÁK*, Pavel VEJL*, Sylva SKUPINOVÁ*, Martina BARDOVÁ*, Jaroslava DOMKÁŘOVÁ**, Lukáš KREUZ*, Pavel KŘENEK*

*KGOZ AF ČZU, **VÚB, Havlíčkův Brod

Souhrn, klíčová slova

Selekce s využitím molekulárních markerů (Marker Assisted Selection - MAS) představuje aktuální trend ve šlechtění rostlin, který umožňuje výrazně urychlit šlechtitelský proces. V tomto příspěvku je prezentována možnost aplikace PCR markeru pro detekci dominantní alely H1 řídící rezistenci bramboru vůči Ro1 patotypu Globodera rostochiensis. Metoda CAPS byla použita pro detekci dominantní alely R1 genu rezistence k Phytophthora infestans.

brambory, Solanum tuberosum, Globodera rostochiensis, Phytophthora infestans, rezistence, DNA, PCR, CAPS.

Summary, keywords

Marker Assisted Selection is a progressive trend of breeding strategy, which can markedly speed up the breeding process. In this paper is represented the possibility of application of PCR marker for detection of dominant allele H 1 controlling resistance of potato against Ro1 pathotype Globodera rostochiensis. The CAPS method was used for detection of dominant allele R1 of resistance against to Phytophthora infestans.

Potato, Solanum tuberosum, Globodera rostochiensis, Phytophthora infestans, resistance, DNA, PCR, CAPS.

Introduction

Marker Assisted Selection represents an effective method of unambiguous detection of some majorgenes controlling studied resistances.

Huijsman (1955) has described dominant controlled resistance against Ro1 pathotype G. rostochiensis - gene H1. Gene origin of this resistance was sub-species S. tuberosum ssp. andigena Hawkes CPCI1673. By means of method RFLP was gene H1 mapped by Gebhardt (1994). In her procedure she has worked with several RFLP probes type cDNA. Niewöhner et al. (1995) have performed controversy of RFLP marker to specific PCR (Polymerase Chain Reaction) marker.

Donator of qualitative resistance against late blight (gene R1) is botanic species Solanum demissum Lindl. Digesting of monomorphic PCR product for detection of dominant allele R1 corresponding with resistance describe Rouppe van der Voort et al. (1998) and De Jong et al. (1997).

Methods

For analysis was taken 90 variants with declared presence or absence of dominant alleles H1 and R1. For isolation of DNA was used kit GenElute Plant Genomic DNA Kit” (Sigma, SRN). For detection of studied alleles were used modified methods according to Niewöhner et al. (1995) and De Jong et al. (1997).

Results - discussion

In all of genotypes, which had declared dominant alleles H1 and R1 was by means of these methods confirmed presence of these alleles. On figures 1 a 2 are shown a typical electrophoretic diagrams that corresponds with Niewöhner et al. (1995) and De Jong et al. (1997) results. By means of PCR or CAPS markers were confirmed the presences of dominant alleles in all of genotypes, which had dominant alleles H1 and R1 declared.

Figure 1: 760bp PCR marker of dominantly allele H1 for resistance against the Ro1 pathotype of G. rostochiensis

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 S

Resistant cultivars: 1 - Accent, 2 - Arnika, 3 - Berber, 4 - Bettina, 5 - Darwina, 6 - Granola, 7 - Gusta, 8 - Hilta, 9 - Ilse, 10 - Indra, 11 - Lyra, 12 - Mira, 13 - Olga, 14 - Ponto, 15 - Tanja, 16 - Ute. Susceptible cultivars: 17 - Ornela, 18 - Tábor, S - leader ëDNA/Eco47I (AvaII)

Figure 2: CAPS marker SPUD237 (digestion of AluI) for detection of resistance against late blight - RI allele

S2 1 2 3 4 5 6 7 8 9 10 S1

Resistant cultivars: 1 - Bionta, 2 - Escort, 3 - Fresco, 4 - Hertha, 6 - Panda, 7 - Premiere, 10 - Ukama. Susceptible cultivars: 5 - Karlena, 8 - Rosara, 9 - Rosella, S1 - leader GeneRulerTM, S2 - leader ëDNA/Eco47I (AvaII)

References

De Jong, W., Forsyth, A., Leister, D., Gebhardt C., Baulcombe, D. C.: Theoretical and Applied Genetics, 95: 246-252, 1997.

Gebhardt, Ch: 71: American Potato Journal, 339-345, 1994.

Huijsman, C. A.: Euphytica, 4: 133-140, 1955.

Niewöhner, J., Salamini, F., Gebhardt, Ch.: Molecular Breeding, 1: 67-78, 1995.

Rouppe van der Voort, J. N. A. M, Lindeman, W., Folkertsma, R., Hutten, R., Overmars, W., van der Vossen, E., Jacobsen, E., Bakker, J.: Theoretical and Applied Genetics 96,: 654 - 661, 1998.

This paper is result of grants NAZV MZe ČR EP9107, FRVŠ 1341/2001. FRVŠ1347/2001,ČZU 21160/1312/213143 and MSM 412100002