Indian Phytopath.

65 (2) : 147-150 (2012)

RESEARCH ARTICLE

Sugarcane genetic resistance against holoparasitic angiosperm Aeginetia pedunculata (Orobanchaceae)

BIKASH RANJAN RAY1* and MRINAL KANTI DASGUPTA2
1 2

Sugarcane Research Station, Bethuadahari 741126, Nadia Department of Plant Protection, Institute of Agriculture, Visva-Bharati, Sriniketan 731 236, Birbhum

ABSTRACT: Twenty eight sugarcane genotypes were screened against the total root parasitic angiosperm Aeginetia pedunculata (Orobanchaceae) by root inoculation with parasite seed. All the entries tested showed parasitization and wilt but variations were observed in respect of first flowering, growing degree days (GDD), area under Aeginetia progress curve (AUAPC) of the parasite and wilting of cane. Breeding of sugarcane by making crosses involving NCo 310 (resistant to A. indica) as either male or female parent resulted in development of genotypes susceptible to A. pedunculata. Key words: Aeginetia pedunculata, resistance breeding, sugarcane

Sugarcane (Saccharum sp., complex hybrid) is an important crop for production of commercial cane sugar (sucrose), molasses (jaggery) or khandseri in India. It is a long duration crop (early = 300 days, mid-late = 360 days), usually one or more ratoon crops are maintained for additional return and its tall dense canopy attracts large number of insect pests and diseases throughout the year and its vegetative propagation allows transmission of most of the diseases for generations. Wilt is a major disease in sugarcane and one of its causal organisms is parasitic angiosperms (Martin et al., 1960). Important parasitic angiosperms of sugarcane are Striga spp. and Centranthera nepalensis D. Don in scrophulariaceae family (obligate root hemiparasite), Aeginetia indica L., A. pedunculata (Roxb.) Wall., A. saccharicola Bakh., Christisonia wightii Elmer and Alectra fluminensis (Vell.) Stearn in Orobanchaceae family (obligate root holoparasite) and Thesium australe R.Br. and Thesium residoides A.W. Hill. in Santalaceae family (facultative root hemiparasite). Striga angustifolia (D.Don) C.J. Saldanha, S. aspera Benth., S. forbesii Benth., S. hermonthica (Del.) Benth., S. latericea Vatke, S. pubiflora Klotzsch infect sugarcane in many countries in sub-Saharan Africa (Atera et al., 2011), S. asiatica (L.) Kuntze infects sugarcane in peninsular India (Chinnusamy et al., 2009) and S. densiflora Benth. infects sugarcane in Bangladesh (Matin et al., 1989), Alectra fluminensis is a minor pest of sugarcane in South America (Parker and Riches, 1993) and Aeginetia spp. affect sugarcane in tropical and sub-tropical Asia. Incidence of A. pedunculata was reported on sugarcane in Myanmar (Subramaniam, 1936) and Bangladesh (Hedayetullah and Saha, 1942). A. pedunculata infected sugarcane in Indonesia (Coert, 1928). Since then Aeginetia spp. were never reported on sugarcane or any other crop. Of late, first major epidemic of A. pedunculata on sugarcane was reported in India in and around a sugar factory zone in West Bengal (Ray and Dasgupta, 2003). It has raised serious threat of further spread and apprehended development of more virulent races. The crop loss in sugarcane due to
*Corresponding author: brray2@yahoo.co.in

infection of the parasite was estimated to 38% in cane yield, 52% in juice brix, 58% in sucrose and 1.89 t/ha in commercial cane sugar (Ray and Dasgupta, 2006). The parasite thrived naturally on a few species of grasses (Poaceae) viz. Cynodon dactylon, Saccharum spontaneum, Sorghum bicolor and Vetiveria zizanoides, adjacent to the infected sugarcane field, they might play a role as collateral hosts in survival and dissemination of the parasite (Ray and Dasgupta, 2009). To control the parasite, the options are: (i) manual weeding soon after appearance (local farmers practice); (ii) 2,4-D Na at 2 kg ai/ha (sugar factory farm managers practice; short-lived success); (iii) resistant varieties (suggestions made; most viable). Large scale replacement of sugarcane varieties with field resistant cv. NCo 310 proved successful in eradicating A. indica in Taiwan (Lo, 1955). Variability of susceptibility to Striga hermonthica (Del.) Benth. was observed in Kenya where among 18 sugarcane clones KEN 83-1228, KEN 83-538 and Co 617 were found relatively tolerant to Striga (Mbogo and Osoro, 1992). Erianthus arundinaceous (Retz.) Jeswiet, which is a source of resistant gene in sugarcane was found susceptible to A. pedunculata (Ray and Dasgupta, 2010). The objectives of the present investigation were: (1) to evaluate the relative tolerance / resistance of elite sugarcane genotypes / varieties to A. pedunculata and (2) to undertake a breeding programme involving NCo 310, a sugarcane variety known to be resistant to A. indica in Taiwan, with other good Indian varieties and to evaluate the existence and inheritance of resistance against A. pedunculata among progenies.

MATERIALS AND METHODS

Screening of sugarcane genotypes for their resistance to A. pedunculata: Single node stalk cuttings of 28 sugarcane varieties/elite genotypes (Table 1) were germinated in polythene packets filled with sugarcane farm

148 Table 1. Characteristics of sugarcane genotypes evaluated against A. pedunculata Sl. Sugarcane Maturity duration ML ML E E ML E E E ML ML E ML E ML E ML ML E E ML E E E ML ML E ML ML Resistance against major diseases Red rot R MR R R MR R R MR R MR MR MS MR MR R R MR R MR MR R MS MR R MR MR R MR Smut R R R R R R R R R R R MR MR R R R R R R R R MR R R R R R S Wilt R MR MR R R R MR MR R R MR S MR R MR R MR R R MR R MS MR MR MR R MR MS

Indian Phytopathology 65 (2) : 147-150 (2012)

No. genotype 1. 2. 3. 4. 5. 6. 7. 8. 9. BO 91 BO 128 BO 140 BO 143 Co 62033 Co 87268 Co 0230 Co 0231 Co 0234

10. Co 0236 11. CoB 99161 12. CoBln 90006 13. CoBln 94063 14. CoLk 92238 15. CoLk 94184 16. CoS 767 17. CoSe 92423 18. CoSe 95436 19. CoSe 96234 20. CoSe 96436 21. CoSe 98021 22. CoSe 00235 23. CoSe 00421 24. CoSe 01232 25. CoSe 01434 26. CoSe 02235 27. UP 1108 28. NCo 310 (Check)

plant spacing of 90cm and 60cm, respectively. Recommended fertilizer dose 200:100:100 (N:P:K) kg/ha, irrigation and intercultural operations were followed and no weeding was done. The experiment was conducted during 2005 and 2006 as plant and ratoon crops, and the varieties were replicated thrice. Data recorded on number of A. pedunculata flowers per sugarcane clump, both in plant and ratoon crops, at approximately weekly intervals starting from anthesis. Date of first flowering was recorded and growing degree days (GDD) for plant and ratoon crop was calculated by cumulating the temperature of each day starting from inoculation to first flowering in respective crop. Area under Aeginetia progress curve (AUAPC) was calculated on the basis of number of A. pedunculata flowers in a clump daily with the help of a MS Excel based AUDPC Calculator Programme. As Aeginetia spp. does not develop any external symptom on sugarcane prior to wilting, a disease rating system was adopted to assess wilt severity according to variety using 1-9 scale similar to Striga rating (Berner et al., 1997) detailed in Table 2. Breeding of sugarcane for resistance to A. pedunculata: A breeding programme was undertaken under National Hybridization Programme of All India Coordinated Research Project on Sugarcane, with the objective of developing A. pedunculata resistant varieties by crossing sugarcane cv. NCo 310 with other sugarcane varieties having records of higher yield, juice quality, agronomic characters and combining ability. The crossing programme was carried out at the National Hybridization Garden of Sugarcane Breeding Institute, Coimbatore during November 2003 and 2004. The treatments were T 1 Co 87272 NCo 310, T2 NCo 310 CoJ 46, T3 CoLk 91238 NCo 310, T4 CoLk 8002 NCo 310, T5 CoLk 8102 NCo 310, T6 Co 1158 NCo 310, T7 BO 91 (Susceptible check) and T8 NCo 310 (Resistant check). The fluff (crossed seed) was received during February and the seeds were sown immediately in soil beds at Sugarcane Research Station, Bethuadahari, West Bengal, India to raise seedlings. Seedlings were transplanted during June to obtain full grown sugarcane plants through vegetative propagation in the following years. A. pedunculata seeds were inoculated to the root zone of these genotypes during May and data recorded on incidence of the parasite and wilting of sugarcane after emergence of its flowers in July. In a separate plot two rows of 3 m length were planted with the same set of genotypes and inoculated with mixed cultures of red rot pathogen Colletotrichum falcatum Went pathotypes Cf 07 and Cf 08 in August by plug method of inoculation (Butler and Khan, 1913) and

Source: Annual Report, All India Coordinated Research Project on Sugarcane 2005-2006; 2006-2007, E = early (300 days), ML = midlate (360 days), R = resistant, MR = moderately resistant, MS = moderately susceptible, S = susceptible

soil and mixed with 5 mg fresh seeds of A. pedunculata. NCo 310 and BO 91 (highly susceptible to A. pedunculata at Plassey, West Bengal, India) were taken as standard check varieties. After sprouting of sugarcane, intact polypacks sets (to avoid underground spread) were planted in an experimental plot at Sugarcane Research station, Bethuadahari, West Bengal, India, with inter-row and inter-

Table 2. Disease rating scale adopted for Aeginetia pedunculata wilt of sugarcane Score

A. pedunculata shoot /
sugarcane clump

Score

Wilted stalk / sugarcane clump (%)

Cumulative score 1 2-3 4-5 6-7 8-9

Disease rating

0 1 2 3 4

0 1-5 6-10 11-20 > 20

1 2 3 4 5

0 1-25 26-50 51-75 76-100

Highly resistant (HR) Resistant (R) Moderately resistant (MR) Susceptible (S) Highly susceptible (HS)

Indian Phytopathology 65 (2) : 147-150 (2012)

Table 3. Genetic resistance in sugarcane genotypes against A. pedunculata wilt Sl. No. Sugarcane genotype First flower (days) 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 26. 27. 28. BO 91 BO 128 BO 140 BO 143 Co 62033 Co 87268 Co 0230 Co 0231 Co 0234 Co 0236 CoB 99161 CoBln 90006 CoBln 94063 CoLk 92238 CoLk 94184 CoS 767 CoSe 92423 CoSe 95436 CoSe 96234 CoSe 96436 CoSe 98021 CoSe 00235 CoSe 00421 CoSe 01232 CoSe 01434 CoSe 02235 UP 1108 NCo 310 (Check) 127 146 0 137 137 174 0 0 122 0 127 0 0 179 132 0 0 0 0 0 0 138 0 0 0 0 0 137 Plant crop First flower (GDD) 2391 2696 0 2566 2566 3068 0 0 2297 0 2391 0 0 3162 2472 0 0 0 0 0 0 2360 0 0 0 0 0 2566 63 269 0 468 300 77 0 0 249 0 770 0 0 14 241 0 0 0 0 0 0 21 0 0 0 0 0 0 AUAPC Disease rating S HS HS HS S S HS MR S MR 0 402 421 407 0 0 358 377 358 402 407 413 402 426 421 407 462 407 407 421 402 358 433 383 524 433 480 433 First flower Ratoon crop First flower (days) 0 6151 6515 6664 0 0 5862 6226 6151 6151 6644 6922 6151 7028 6515 6664 7724 6664 6664 6515 6151 5862 6625 6336 8778 6625 8061 6625 AUAPC (GDD) 0 51 83 74 3 0 1061 100 253 96 102 189 110 57 74 688 30 93 380 6 144 9 112 90 44 89 56 139

149

Disease rating S S S HS S HS S HS MR HS MR MR HS S S HS S S MR S S S S MR S

Table 4. Breeding of sugarcane for development of A. pedunculata resistant cultivar Treatment T1 T2 T3 T4 T5 T6 T7 T8 Cross (Male Female) Co 87272 NCo 310 NCo 310 CoJ 46 CoLk 91238 NCo 310 CoLk 8002 NCo 310 CoLk 8102 NCo 310 Co 1158 NCo 310 BO 91 (Susceptible check) NCo 310 (Resistant check) No. of clone 30 26 4 16 7 25 1 1 Brix (%) 16.9 18.6 17.9 16.9 18.0 17.8 19.5 16.7 Red rot rating MS MS S MS MR MR MR MR

A. pedunculata wilt rating

S S HS HS S S S S

data recorded after 60 days in 0-9 scale by splitting open the cane longitudinally along the point of inoculation.

RESULTS AND DISCUSSION

First flowers of A. pedunculata appeared in the root zones of sugarcane variety BO 91, BO 128, BO 143, Co 62033, Co 87268, Co 0234, CoB 99161, CoLk 91238, CoSe 94184

and CoSe 00235 during July to September in both the years, approximately 3-5 months after inoculation and one month after a heavy downpour but in case of other varieties, flowering occurred in the following year. The morphological features of the flowers resembled with that usually occur in the sugar factory area. Infected sugarcane clumps invariably wilted and dried after completion of flowering of A. pedunculata. During first year, total wilting of clumps were

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Indian Phytopathology 65 (2) : 147-150 (2012)

observed in BO 91 and Co 87268 and partial wilting was observed in BO 128, BO 143, Co 62033, CoB 99161, CoSe 94184 and CoSe 00235, whereas, in rest of the varieties wilting occurred in the second year. Only one or two new sugarcane shoots came out in the third year but no flower of A. pedunculata was observed. Though, all the sugarcane genotypes were resistant or moderately resistant to major fungal diseases such as red rot, wilt and smut, none of the entries showed resistance to A. pedunculata, even cv. NCo 310, which was widely used as resistant variety to eradicate A. indica in Taiwan and used as check variety in this experiment turned out to be highly susceptible to A. pedunculata (Table 3). The screening procedure was simple and reasonably effective in the field. By nature, the parasite does not spread beyond the experimental area and caused no threat to the farm cultivation. The sugarcane entries showed significant variability among themselves in respect of first flowering, duration of flowering and flower (Table 1). The AUAPC data represented the number of flower and duration of flowering in A. pedunculata, which varied widely according to host (Table 3). The susceptibility of NCo 310 to A. pedunculata further confirmed our earlier claim that these two species of Aeginetia are distinct in respect of their host range and that A. pedunculata is more virulent than A. indica. In fine, no sugarcane genotype was identified as tolerant or resistant to A. pedunculata and therefore, no variety could be included in IPM. Although, there is no A. pedunculata resistant cultivar in the genetic stock the variation in susceptibility and selective host range of A. pedunculata among Poaceae indicate that occurring of genetic resistance is possible. Primary objective of the resistant breeding programme was to incorporate resistant gene into the established sugarcane cultivars because genetic resistance is the most viable and eco-friendly option for management of parasitic angiosperms. The only known gene source for resistance against A. indica was NCo 310 and considering A. indica and A. pedunculata are closely related species, 110 clonal lines were developed through the crosses between NCo 310 and any one of the following varieties as male or female parent Co 87272, CoJ 46, CoLk 91238, CoLk 8002, CoLk 8102 and Co 1158. Though, these genotypes were good yielder of sucrose (16.9%-18.6%) and moderately resistant to red rot disease, they were susceptible to A. pedunculata (Table 4). The result was expected because in a later experiment NCo 310 was proved to be susceptible to A. pedunculata . This differential susceptibility between A. indica and A. pedunculata to NCo 310 might be due to genetic variance between the parasites or might be due to clonal variance of host or its environment.

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Atera, E.A., Itoh, K. and Onyango, J.C. (2011). Evaluation of ecologies and severity of Striga weed on rice in sub-Saharan Africa. Agriculture and Biology Journal of North America 2(5): 752-760. Berner D.K., Winslow, M.D., Awad, A.E., Cardwell, K.F., Mohan Raj, D.R. and Kim, S.K. (1997). Striga Research Methods A Mannual. International Institute of Tropical Agriculture, Ibadan, Nigeria pp. 59-60. Butler, E.J. and Khan, H.A. (1913). Red rot of sugarcane. Memoirs of Department of Agriculture, India. Botanical Series 6: 151-178. Chinnusamy, C., Prabhakaran, N.K. and Rathika, S.. (2009). Ecobiological quantification and integrated management of parasitic weed Striga asiatica in sugarcane ( Saccharum officinarum) planted in alfasol of southern peninsular India. Poster, 10th World Congress on Parasitic Plants, Kusadasi, Turkey, June 8-12, 2009. Coert, J.H. (1928). Aeginetia species, a root parasite of sugarcane. Sugar News 9: 367-375. Hedayetullah, S. and Saha, J.C. (1942). A new phanerogamic parasite of sugarcane in Bengal. Curr. Sci. 11(3): 109-110. Kusano, S. (1908). Further studies on Aeginetia indica . Bulletin of the College of Agriculture, Tokyo Imperial University, Japan. 8: 59-76. Lo, T.T. (1950). A report on sugarcane diseases in Taiwan. Proceedings of ISSCT Congress 7: 452-456. Lo, T.T. (1955). N:Co 310, highly resistant to the root parasite bunga (Aeginetia indica). Taiwan Sugar 2(4): 18-20. Lopez, M.E. and Barile, R.L. (1964). The effect of different herbicides in the control of Aeginetia indica Roxb. Philippine Sugar Institute Quarterly 10(1): 19-31. Martin, J.P., Abbott, E.V. and Hughes, C.G. (1960). Sugarcane Diseases of the World, Vol. I. Elsevier, Amsterdam. pp 485490. Matin, M.A., Islam, N., Gaffer, M.A. and Rahman, A.B.M.M. (1989). Association of Striga densiflora with sugarcane. Bangladesh Journal of Sugarcane 11: 52-63. Mbogo, J.O. and Osoro, M.O. (1992). The effect of Striga hermonthica on sugarcane, 1. Reacton of sugarcane clones to Striga hermonthica infection in Kenya. Proceedings of the Annual Congress (66 th) South African Sugar Technologists Association, pp. 114-115. Parker, C. and Riches, C.R. (1993). Parasitic Weeds of the World: Biology and Control. CAB International, Wallingford, UK. p. 332. Ray, B.R. and Dasgupta, M.K. (2003). First report of Aeginetia pedunculata causing sugarcane wilt in India. Haustorium 44: 2-3. Ray, B.R. and Dasgupta, M.K. (2006). Sugarcane crop loss due to wilt caused by parasitic angiosperm Aeginetia pedunculata (Roxb.) Wall. (Orobanchaceae). J. Mycol. Plant Pathol. 36 (1): 31-34. Ray, B.R. and Dasgupta, M.K. (2009). Three newly recorded natural hosts of Aeginetia pedunculata (Roxb.) Wall. (Orobanchaceae). J. Mycol. Plant Pathol. 39 (1):163-165. Ray, B.R. and Dasgupta, M.K. (2010). Erianthus arundinaceus: a new host of Aeginetia pedunculata. J. Mycol. Plant Pathol. 40 (2): 283-286. Subramaniam, L.S. (1936). Diseases of Sugarcane and Methods for their Control. Imperial Council of Agricultural Research, Miscellaneous Bulletin No. 10. pp. 25-26. Received for publication: September 18, 2011 Accepted for publication: April 16, 2012

ACKNOWLEDGEMENTS
Authors are grateful to Dr. Kunal Mondal, Plant Pathologist, National Research Centre for Medicinal & Aromatic Plants, Boriavi, Anand, Gujarat, India, for providing MS Excel based AUDPC Calculator which he had designed. They are also grateful to the Director, Sugarcane Breeding Institute, Coimbatore for supply of sugarcane variety NCo 310 and for undertaking crossing programme comprising NCo 310.