Functional stacking of three resistance genes against Phytophthora infestans in potato - PubMed (original) (raw)

Functional stacking of three resistance genes against Phytophthora infestans in potato

Suxian Zhu et al. Transgenic Res. 2012 Feb.

Abstract

Functional stacking of broad spectrum resistance (R) genes could potentially be an effective strategy for more durable disease resistance, for example, to potato late blight caused by Phytophthora infestans (Pi). For this reason, three broad spectrum potato R genes (Rpi), Rpi-sto1 (Solanum stoloniferum), Rpi-vnt1.1 (S. venturii) and Rpi-blb3 (S. bulbocastanum) were selected, combined into a single binary vector pBINPLUS and transformed into the susceptible cultivar Desiree. Among the 550 kanamycin resistant regenerants, 28 were further investigated by gene specific PCRs. All regenerants were positive for the nptII gene and 23 of them contained the three Rpi genes, referred to as triple Rpi gene transformants. Detached leaf assay and agro-infiltration of avirulence (Avr) genes showed that the 23 triple Rpi gene transformants were resistant to the selected isolates and showed HR with the three Avr effectors indicating functional stacking of all the three Rpi genes. It is concluded that Avr genes, corresponding to the R genes to be stacked, must be available in order to assay for functionality of each stack component. No indications were found for silencing or any other negative effects affecting the function of the inserted Rpi genes. The resistance spectrum of these 23 triple Rpi gene transformants was, as expected, a sum of the spectra from the three individual Rpi genes. This is the first example of a one-step approach for the simultaneous domestication of three natural R genes against a single disease by genetic transformation.

PubMed Disclaimer

Figures

Fig. 1

Fig. 1

Graphical view of binary vector pBINPLUS:_Rpi_-blb3:_Rpi_-vnt1.1:_Rpi_-sto1. Restriction enzyme cleavage sites, used for cloning of the Rpi genes into pBINPLUS have been indicated

Fig. 2

Fig. 2

Agro-infiltration of Avr effectors in triple Rpi gene regenerant A14 Z-22 using different densities of inoculum. Clear HR reactions were observed at all densities for Avr2, IpiO1 and Avrvnt1. Co-infiltration of R3a/Avr3a, and vnt1.1/_Avrvnt_-1 served as a positive controls. Empty vector pGRAB, and Agrobacterium strain AGL1 were negative controls. a Showed the infiltration with Avr2 and IpiO1. b Showed the infiltration with Avrvnt1

Similar articles

Cited by

References

    1. Agrawal PK, Kohli A, Twyman RM, Christou P. Transformation of plants with multiple cassettes generates simple transgene integration patterns and high expression levels. Mol Breed. 2005;16:247–260. doi: 10.1007/s11032-005-0239-5. - DOI
    1. Altpeter F, Baisakh N, Beachy R, Bock R, Capell T, Christou P, Daniell H, Datta K, Datta S, Dix PJ, Fauquet C, Huang N, Kohli A, Mooibroek H, Nicholson L, Nguyen TT, Nugent G, Raemakers K, Romano A, Somers DA, Stoger E, Taylor N, Visser RGF. Particle bombardment and the genetic enhancement of crops: myths and realities. Mol Breed. 2005;15:305–327. doi: 10.1007/s11032-004-8001-y. - DOI
    1. Anand A, Zhou T, Trick HN, Gill BS, Bockus WW, Muthukrishnan S. Greenhouse and field testing of transgenic wheat plants stably expressing genes for thaumatin-like protein, chitinase and glucanase against Fusarium graminearum. J Exp Bot. 2003;54:1101–1111. doi: 10.1093/jxb/erg110. - DOI - PubMed
    1. Andreu AB, Caldiz DO, Forbes GA. Phenotypic expression of resistance to Phytophthora infestans in processing potatoes in Argentina. Am J Pot Res. 2010;87:177–187. doi: 10.1007/s12230-009-9121-z. - DOI
    1. Armstrong MR, Whisson SC, Pritchard L, Bos JIB, Venter E, Avrova AO, Rehmany AP, Bohme U, Brooks K, Cherevach I, Hamlin N, White B, Fraser A, Lord A, Quail MA, Churcher C, Hall N, Berriman M, Huang SW, Kamoun S, Beynon JL, Birch PRJ. An ancestral oomycete locus contains late blight avirulence gene Avr3a, encoding a protein that is recognized in the host cytoplasm. PNAS. 2005;102:7766–7771. doi: 10.1073/pnas.0500113102. - DOI - PMC - PubMed

Publication types

MeSH terms

LinkOut - more resources