Sunflower broomrape (Orobanche cumana Wallr.) is a parasitic plant which has a significant negative impact on seed
yield. The parasite is spread in large areas of Europe, Asia and it has identified recently, in North Africa. Breeding for
resistance is regarded as the most effective, feasible and environmentally friendly solution to control sunflower
broomrape. However, breeding for resistance is challenging as new races of the parasite have evolved. The use of
resistant hybrids of monogenic resistance type, is followed by the appearance of new more virulent races that overcome
the existing resistance genes. So, it is necessary to develop sunflower hybrids which can accumulate qualitative and
quantitative resistance in a single one, in order to have a durable resistance. Among this, by developing Clearfield
Production System in sunflower it could have an important control strategy and complemented the genetic resistance
against the parasite

Anton, Gabriel

Bran, Alexandru

Dan, Mihaela

Florea, Andreea-Mihaela

Lipșa, Florin-Daniel

Pacureanu, Maria Joita

Poaleungi Prodan, Tudorita

Risnoveanu, Luxita

Sava, Elisabeta

Ulea, Eugen

Poalelungi Prodan, Tudorița

Joița Păcureanu, Maria

Rîșnoveanu, Luxița

Anton, Florin-Gabriel

ALSE Repository

Lucrări Ştiinţifice – vol. 63(2)/2020, seria Agronomie  75   BROOMRAPE (OROBANCHE CUMANA WALLR.) CONTROL BY DEVELOPING GENETIC RESISTANT GENOTYPES IN SUNFLOWER   Tudorita (POALEUNGI) PRODAN1, Maria JOITA PACUREANU2, Luxita RISNOVEANU4,5,  Mihaela DAN2, Gabriel ANTON2,3, Alexandru BRAN6, Elisabeta SAVA6, Florin-Daniel LIPȘA1, Andreea-Mihaela FLOREA1, Eugen ULEA1  e-mail: dorina.tudoritaprodan@yahoo.com   Abstract  Sunflower broomrape (Orobanche cumana Wallr.) is a parasitic plant which has a significant negative impact on seed yield. The parasite is spread in large areas of Europe, Asia and it has identified recently, in North Africa. Breeding for resistance is regarded as the most effective, feasible and environmentally friendly solution to control sunflower broomrape. However, breeding for resistance is challenging as new races of the parasite have evolved. The use of resistant hybrids of monogenic resistance type, is followed by the appearance of new more virulent races that overcome the existing resistance genes. So, it is necessary to develop sunflower hybrids which can accumulate qualitative and quantitative resistance in a single one, in order to have a durable resistance. Among this, by developing Clearfield Production System in sunflower it could have an important control strategy and complemented the genetic resistance against the parasite.  Key words: sunflower broomrape, genetic control, qualitative resistance, quantitative resistance, herbicides resistance                                                    1 “Ion Ionescu de la Brad” University of Agricultural Sciences and Veterinary Medicines, Iasi 2 National Agricultural Research and Development Institute, Fundulea 3 University of Agronomic Sciences and Veterinary Medicine, Bucharest 4 Agricultural Research Development Station, Braila 5 „Dunarea de Jos” University of Galati, Engineering and Agronomy Faculty of Braila ⁶ The State Institute for Variety Testing and Registration The parasitic weed Orobanche cumana (sunflower broomrape) is an obligatory and non-photosynthetic root parasitic plant of the sunflower (Helianthus annuus L.) and is a substantial threat in Europe, especially in countries around the Black Sea and in Spain (Molinero-Ruiz L. et al, 2013, Louarn J. et al, 2016). Under favourable conditions, it infects the roots of sunflower plants and connects to the vascular tissue, thus depleting the nutrients and affecting host growth and yield (Heide-Jorgensen H., 2008; Molinero-Ruiz L. et al, 2015). Broomrape seeds are very small and individual plants can produce an impressive number of seeds that remain viable in the soil for up to 20 years. They are widely disseminated by water, wind, animals, humans, machinery, or though attachment to sunflower seeds (Parker C., 2013).  A major difficulty for the breeders is the fast development of new races of the parasite, which overcome the resistance of sunflower genotypes. To the present day, more than seven races of sunflower broomrape have been identified (Kaya Y., 2014). Vranceanu A.V. et al (1980) identified five races of O. cumana, designated as A, B, C, D and E. Later on, more virulent race F was identified in Spain (Alonso L.C. et al, 1996; Molinero-Ruiz L. et al, 2008, Martin-Sanz et al 2016), Romania (Păcureanu-Joita M. et al, 1998), Turkey (Kaya Y. et al, 2004) and some other counties. Presence of more virulent broomrape races, designated G and H, has been also reported (Shindrova P. and Penchev E., 2012, Antonova L. et al, 2014, Kaya Y., 2014). Current racial situation of broomrape in the main infested areas is unclear, since there is a lack of information on whether races under the same name reported in different countries are the same or differ in terms of virulence (Fernández-Martínez J. et al, 2012; Molinero-Ruiz L. et al, 2015, Martin-Sanz A. et al, 2016). Breeding for genetic resistance appears to be the most appropriate and reliable measure to control the parasite. The changes in broomrape race composition have forced sunflower breeders to continuously search for resistance genes to new races and study their genetic control.  Universitatea de Ştiinţe Agricole şi Medicină Veterinară Iaşi  76 The objective of this study was to identify sources of resistance to the parasite and to develop genetic resistant genotypes to broomrape.  MATERIAL AND METHOD  Six cultivated sunflower inbred lines and two sunflower wild species have been introduced in crossing for obtaining interspecific hybrids, in order to create sunflower populations which will be used for releasing inbred lines with high resistance to broomrape (Orobanche cumana).  These hybrids as well as the parental forms (cultivated and wild) have been studied for resistance to the parasite broomrape. There have been analyzed the number of sunflower heads and number of seeds/head, for each crossing. The crossing between cultivated and wild sunflower was made by emasculation in cultivated inbred lines and making pollination with wild species pollen, as well as making emasculation in wild forms and pollination with pollen of cultivated ones.  The populations which were obtained from interspecific hybrids have been tested for resistance to broomrape. The resistance to broomrape parasite was made in natural and artificial infestation conditions. The testing in the artificial infestation was made in glass house, in pots of 5 liters capacity, having inside a mixture of soil and sand (3/1) as well as broomrape seeds (1 g/pot). It has been collected broomrape from areas very high infested with the parasite. In natural infestation, the testing was made in two locations situated in areas Constanta and Tulcea, with different virulence of broomrape populations.  RESULTS AND DISCUSSIONS  The results regarding the crosses between wild and cultivated sunflower are presented in table 1. In case of using cultivated sunflower as pollen receptor, the number of seeds/head is high, comparing with case of using the wild sunflower as pollen receptor, when the number of seeds/head is very low. In this case the number of heads is high, taking into consideration that the wild sunflower is high branched, so, there are many small heads. Table 1 Results regarding the crosses between wild and cultivated sunflower Helianthus annuus H. tuberosus H. agrestis LC 1001 LC 985 LC 1015 LC 1066 LC 1085 LC 1088  Hybridization: number of heads and seed/heads LC 1001 B 3/640 3/250       LC 985 B 3/520 3/120       LC 1015 B 3/390 3/360       LC 1066 C 3/150 3/150       LC 1085 C 3/280 3/250       LC 1088 C 3/130 2/110       Helianthus tuberosus   25/15 25/7 25/5 25/14 25/23 25/12  In figure 1 are presented the results regarding the resistance to broomrape for sunflower populations obtained from interspecific hybrids. Comparing the results obtained in two locations situated in Tulcea and Constanta areas, these are showing that some populations are full resistant in Tulcea area, having a low attack degree in Constanta area.  The sunflower differential line, for the race E which was used as check for sensitivity, has a high infestation degree, in both locations. This it means that in these locations the parasite has developed races more virulent.  Figure 1 Results regarding the resistance of some sunflower populations obtained from interspecific hybrids, to the broomrape parasite, in the natural infestation conditions, in two areas in Romania Lucrări Ştiinţifice – vol. 63(2)/2020, seria Agronomie  77 In figure 2 is presented the scheme used for introducing the genes for resistance to the parasite broomrape, in the valuable sunflower inbred lines. In this process of the genes transferring it needs a number of 3-4 backcross generations, followed by 1-2 generation of self-pollination.   Figure 2 - Scheme used for the genes for resistance transferring, in the valuable inbred lines  In figure 3 are presented the results regarding the behavior of some lines in different generations of the genes transfer, using many broomrape populations, from the most infested areas in southeastern Romania.  The testing was done in the artificial infestation conditions. It can be seen that for some lines, the resistance is increasing after each generation of selection, taking into consideration that it is made one generation per year.  Figure 3 Results regarding the behavior of sunflower lines, in different generations of selection, in two years  Universitatea de Ştiinţe Agricole şi Medicină Veterinară Iaşi  78 CONCLUSIONS  The broomrape parasite has become very dangerous for sunflower crop in almost all areas cultivated with sunflower in Europe. It is of a great importance to identify sources of resistance to the new races of broomrape. For this, the sunflower wild species are very important, they being the best source of genes for resistance. Some sunflower populations obtained by crossing sunflower wild species with cultivated genotypes have good resistance to the broomrape races which are spread in the most important areas cultivated with sunflower in Romania. Using a scheme efficient for transferring the genes for resistance to broomrape parasite, in some valuable sunflower inbred lines, there has been obtained high resistant genotypes  REFERENCES  Alonso L.C., Fernandez-Escobar J., Lopez G., Sallago F., 1996 - New highly virulent sunflower broomrape (Orobanche cernua Loefl.) pathotypes in Spain. In Proceedings of 6th International Parasitic Weed Symposium, Seville, Spain, 3-6 June 1996. Seville, Spain: Direccion General de Investigacion Agraria, Consejería de Agricultura y Pesca: 639-644. Fernández-Martínez J., Velasco L., Pérez-Vich B., 2012 - Progress in research on breeding for resistance to broomrape. In Proceedings of 18th International Sunflower Conference, Mar del Plata, Argentina, Feb 27-March 1 2012. Paris, France: International Sunflower Association: 1111-1114. Heide-Jorgensen H., 2008 - Parasitic flowering plants. The Netheralnds: Konin-klijke Brill NV. Kaya Y., Evci G., Pekcan V., Gucer T., 2004 - Determining new broomrape-infested areas, resistant lines and hybrids in Trakya region of Turkey. Helia 27: 211-218. Kaya Y., 2014 - Sunflower. A. Pratap. (Ed) Alien Gene Transfer in Crop Plants, Vol. 2. Springer    Press. 281-315. Kaya Y., 2014 - Current situation of sunflower broomrape around the world. Proceedings of 3rd International Symposium on Broomrape (Orobanche spp.) in Sunflower, Cordoba, Spain, Int Sunflower Assoc, Paris, France: 9-18. Louarn J., Boniface M. C., Pouilly N., Velasco L., Pérez-Vich B., Vincourt P., Muños S., 2016 -  Sunflower resistance to broomrape (Orobanche cumana) is controlled by specific QTLs for different parasitism stages. Front. Plant Sci. 7: 590.  Martín-Sanz A., Malek J,, Fernández-Martínez J.M., Pérez-Vich B., Velasco L., 2016 - Increased Virulence in Sunflower Broomrape (Orobanche cumana Wallr.) Populations from Southern Spain Is Associated with Greater Genetic Diversity. Front. Plant Sci. 7:589. Molinero-Ruiz M.L., Pérez-Vich B., Pineda-Martos R., Melero-Vara J.M., 2008 - Indigenous highly virulent accessions of the sunflower root parasitic weed Orobanche cumana. Weed Res. 48: 169-178. Molinero-Ruiz L., García-Carneros A.B., Collado-Romero M., Raranciuc S., Domínguez J., Melero-Vara J.M., 2013 - Pathogenic and molecular diversity in highly virulent populations of the parasitic weed Orobanche cumana (sunflower broomrape) from Europe. Weed Res. 54: 87–96. Molinero-Ruiz L., Delavault P., Pérez-Vich B., Păcureanu-Joita M., Bulos M., Altieri E., Dominguez J., 2015 - History of the race structure of Orobanche cumana and the breeding of sunflower for resistance to this parasitic weed: A review. Spanish J. Agric. Res. 13:4.  Pacureanu-Joita M., Vranceanu A.V., Seare G., Marinescu A., Sandu I., 1998 - The evaluation of the parasite-host interaction in the system Helianthus annuus L. – Orobanche cumana Wallr. in Romania. In: Proc. 2nd Balkan Symposium on Field Crops, Novi Sad, Yugoslavia: 153-158. Parker C., 2013 - The Parasitic Weeds of the Orobanchaceae, In Parasitic Orobanchaceae, Springer, Berlin, Germany: 313-344 Shindrova P., Penchev E., 2012 - Race composition and distribution of broomrape (Orobanche cumana Wallr.) in Bulgaria during 2007-2011. Helia 35: 87-93. Skoric D., 2012. - Sunflower breeding. In: Skoric, D. (editor), Sunflower genetics and breeding. Serbian Academy of Sciences and Arts, Novi Sad, Serbia, pp. 165-354. Vranceanu A.V., Tudor V. A., Stoenescu F.M., Pirvu N., 1980 - Virulence groups of Orobanche cumana Wallr. differential hosts and resistance sources and genes in sunflower. In: Proc 9th Int Sunfl Conf, Torremolinos, Spain. Int Sunfl Assoc Paris, France: 74-80.  

Broomrape (Orobanche cumana wallr.) control, by developing genetic resistant genotypes in sunflower

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Broomrape (Orobanche cumana wallr.) control, by developing genetic resistant genotypes in sunflower

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