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Volume 3 No 1 January 2013

A Review on Current Research Advancement on Wilt Disease of Psidium Guajava L. with Special Reference to Management paper subtitle: Guava wilt- occurrence, causes and cuers

Dr. Nidhi Didwania Department of Biotechnology Faculty of Engineering and Technology Manav Rachna International University Faridabad, Haryana, India E-mail: nidhididwania77@gmail.com

Neha Katyal Department of Biotechnology Faculty of Engineering and Technology Manav Rachna International University Faridabad, Haryana, India E-mail: katyneha@gmail.com

Deepti Sadana Department of Biotechnology Faculty of Engineering and Technology Manav Rachna International University Faridabad, Haryana, India E-mail: gaganchashaki@yahoo.com Abstract— Guava (Psidium guajava L., family Myrtaceae), ‘‘poor man’s fruit’’ or ‘‘apple of tropics’’, an important fruit crop of several tropical and sub-tropical countries, is facing several agronomic and horticultural problems such as susceptibility to many pathogens, particularly guava wilting (caused by Fusarium solani, F. longipes, F. moniliforme [Gibberella fujikuroi], F. oxysporum f.sp. psidii, Macrophomina phaseolina and Rhizoctonia sp.), low fruit growth, short shelf life of fruits, high seed content, and stress sensitivity.

In India, it is grown in almost all of the states. It is a hardy crop and is cultivated successfully even in neglected soils. Wilt of guava is one of the most important disease of guava especially in India and loss due to this disease is substantial [3]. The disease was first reported by Das Gupta and Rai from the orchards of Lucknow in India [4]. Wilt is the most destructive disease of guava and causes 5– 60% loss in guava production in India [5]. Assessment of loss around Lucknow revealed that losses vary from 5-60 per cent and above 5 year old guava plants are more prone to wilt incidence [6]. Although, more than five decades have been passed since its first report, limited progress has been made in the understanding of the disease.

This review outlines the recent findings made to control the disease severity, including those by cultural, chemical, biological and integrated methods. The modes of infection, causal organisms, and environmental and chemical factors that guide the development of the disease have also been emphasized. Keywords: guava wilt, Psidium oxysporum f. sp. psidii, management

guajava,

II. SYMPTOMATOLOGY Wilt of guava plants showing similar symptoms has been reported in many parts of India [7]. Some trees affected linger on even upto 252days and then die. The variations in the symptoms were also studied during different time of the year. They noticed yellowing of the leaves with interveinal chlorosis during the month of August, which drop even with the slight shaking of the plants. During September general drooping of the leaves take place. During October complete wilting of the plants are seen with almost dried leaves and small dried black fruits hanging on the branch. Few plants also show partial wilting, which is a very common symptom of wilt in guava. It is also recorded that some plants show wilting of variable degree (leaf yellowing, drooping of leaves, drying of terminal branches or partial wilting) during different months but later escape/resist wilting. These plants start recovering from December onwards. It was recorded that

Fusarium

I. INTRODUCTION Guava is considered as one of the exquisite, nutritionally valuable and remunerative crops. Guava fruit contains 2–5 times more vitamin C than orange and is also good source of calcium, phosphorus, and iron. High concentration of pectin in guava fruit may play a significant role in the reduction of cholesterol and thereby decrease the risk of cardiovascular disease [1]. Traditionally, different parts of plants, i.e. fruits, leaves, roots, and bark are used in the treatment of gastroenteritis, diarrheoa, and dysentery [2]. Despite these advantages, there are a number of problems that affect guava production. 8


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out of total wilting plants, around 17 % plants, which initially shows some symptoms of wilting, ultimately escape/resist wilting [8].

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Although several pathogens have been reported for the cause of wilt in guava by different workers but Fusarium oxysporum f. sp. psidii and Fusarium solani were found to be most important pathogen associated with the wilt disease of guava in India [17].

The pathogen attacks young as well as old fruit bearing trees but older trees are more prone to the disease. New seedlings and grafts also show disease symptoms. There are mainly two types of symptoms i. e. slow wilt (where plant takes several months or even a year or two to wilt after the appearance of initial symptoms) and sudden wilt (where plant takes 15 days to one month to wilt after the appearance of initial symptoms).

IV. EPIDEMIOLOGY There is severe incidence of wilt in alkaline soils at pH ranging from 7.5 to 9.0 [18] or in lateritic soils at pH 6.5 [19]. More disease is reported in clay loam and sandy loam compared to heavy soil types [20]. Infected guava plants start showing sign of wilting right with the onset of rainy season in August with maximum number dying in September and October. Soon after rainy season, in September-October, the entire tree dries up

III. CAUSAL ORGANISM The exact cause of the disease is still not fully understood but the pathogens viz. Fusarium oxysporum f. sp. psidii (Prasad, Mehta & Lal), F. solani (Mart.) App. & Wollenw., Macrophomina phaeseoli(Maubl.)Ashby, Rhizoctonia bataticola (Taub.) Bulter, Cephlosporium sp. and Gliocladium roseum (Link ex Fr.) and various pathogens are reported by different workers may be the incitant of the disease. Prior to 1941, wilt was considered to be caused by Cephalosporium sp. [9]. Later, it was attributed that wilt is due to Fusarium oxysporum (Fr.) Schl. and the name Fusarium oxysporum (Fr.) Schl. f.sp. psidii Prasad, Mehta and Lal was proposed [10]. In a study Misra and Pandey and reported that Gliocladium roseum as a most potent pathogen, which reproduces symptom of wilt on artificial inoculation [11].

Guava seedlings are more susceptible to F. solani as well as F. oxysporum f.sp. psidii than older plants of 3 years age. On the other hand researchers reported that F. solani could infect guava plants from 1-month-old to more than 4 years old. A common practice followed in West Bengal that the growers harvest the fruits upto a 6-year-old plants. They then discard the old plants, take new plantation and their escape the wilt incidence. V. VARIETAL REACTION Varieties, white guava No. 6229, Clone.32-12, Webber and Popeno from Florida (USA), Hart and Rolf from Florida but acclimatized at Allahabad, Riverside and Rolf from California (USA), Safeda from Sri Lanka, Banarasi (Andhra strain), Dholka, Sindh and Nasik (Bombay strain) were reported tolerant to wilt disease. Out of 47 varieties of guava screened, the varieties Supreme Clone 32-12, Webber, Popeno, Hart and Rolfs from Florida (USA) were resistant but the varieties Allahabad, Safeda from Ceylon, Banarsi (Andra strain), Dholka (Sindh) and Nasik (Mombay) from India were found as the most tolerant to the disease [21]. The variety (from Baruipur) was fairly resistant against F. solani f.sp. psidii as well as resistant from combined infection by F.solani f.sp. psidii and Macrophomina phaseoli. Allahabad Safeda had fair resistance against combined infection with F. solani f.sp. psidii and M. phaseoli. As far as host resistance was concerned, 10 red-fleshed and 15 white fleshed guava cultivars were screened and it was found that only red fleshed cultivar were infected by F. solani f.sp. psidii [22]. Of the 15 white fleshed cultivars, Lucknow-49 was free from the disease and in Allahabad Safeda; the disease incidence was only 4 percent whereas Kerala and Behat Coconut suffer heavily (33%). None of the other 7 Psidium spp., P. araca, P. cattleianum var. lucidum, P. corecium, P. cujavillus, P. guineese, P. friedrichsthalianum was found affected. Among the cultivars tested, one (Banarasi) was resistant and two (Allahabad Safeda and Seedless) were moderately resistant. Six cultivars were moderately susceptible [23].

A survey conducted in major guava-growing areas in Uttar Pradesh, India during 2001-2003 showed the association of F. oxysporum f. sp. psidii and Verticillium sp. with wilted trees [12]. Isolation and trials on pathogenicity proved to be Verticillium albo-atrum, this being the first report of V. albo-atrum, causing wilt in guava. The wilt was later reported to be caused by Botyodiplodia theobromae in Midnapur (West Bengal) [13] and due to Gliocladium roseum in association with Sclerotium rolfsii and Rhizoctonia solani [14]. Interaction between or among the pathogens may happen during the course of infection. The exact cause of disease is not fully understood and it is recognized as disease complex caused by more than one organism and possibilities not to be ruled out [15]. A survey of nematode and wilt problems of guava (Psidium guajava) was carried out in the Allahabad region and its adjacent areas in Uttar Pradesh. India [16]. Tylenchulus semipenetrans, Xiphinema sp., Longidorus sp. Tylenchorhynchus brassicae, Hoplolaimus indicus [Basirolaimus indicus], Helicotylenchus indicus, Ditylenchus dipsaci and Hemicriconemoides sp. were the predominant phytonematodes. Fusarium oxysporum, F. solani, F. equisetii, F. acuminatum [Gibberella acuminate] F. moniliforme [G. fujikuroi], F. semitectum [F. pallidoroseum] and F. fusarioides [F. chlamydosporum] were the predominant wilt-causing fungi.

VI. PHYSIOLOGICAL STUDIES 9


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Physiology of the disease of guava decline was described by several workers. The cultural and physiological (temperature and pH) characters of F. oxysporum f. sp. psidii and F. solani were studied and the data revealed that maximum mycelial growth was obtained in potato dextrose agar as semi-solid media i.e. 78.00 mm for F. oxysporum f. sp. psidii; 73.83 mm for F. solani, while malt extract broth as liquid broth media i.e. 1 385 mg mycelia for F. oxysporum f. sp. psidii; 1491 mg for F. solani. Maximum sporulation was recorded in oatmeal agar and mycological broth. The optimum temperature and pH for growth of both Fusarium spp. isolates was 28°C and 5.5. The isolates differed in their colony growth; mycelial mass, macro-conidia, and micro-conidia produced. These variations were characters of each of the isolates with respect to cultural and physiological characters [24].

WP the radial growth of Fusarium oxysporum f. sp. psidii was significantly less. This might be due to carbendazim 50% WP is a systemic fungicide which is effective against Fusarium oxysporum f. sp. psidii [29].

VII. DISEASE MANAGEMENT To achieve a meaningful management of the pathogen and a substantial degree of disease control, all the four components of disease pyramid are to be managed. This goal can be achieved by the integration of methods directed against the causal agent, in favour of the host and for modification of the environment. The package of practices consisting of a combination of cultural, biological, chemical methods and host resistance help in reducing the diseases.

C. Biological control Being the soil borne nature of wilt pathogen, it is unpractical to control it with any chemical. The effects of chemicals are also hazardous for the soil and environment, moreover when the effect of chemicals diminishes, the pathogen become more virulent and aggressive. Hence, considering the above facts, it was considered more desirable to use the bio-agents for the control the wilt disease.

B. Cultural practices Wilt could be controlled by proper sanitation in the orchard [30]. Wilted trees should be uprooted, burnt and trench should be dug around the tree trunk. Symptoms of the disease do not appear under green manuring and the disease development is less when organic sources of nitrogen are used [31]. Soil solarization with 30 fm transparent polyethylene sheet during May-June have been suggested for the control of wilt pathogens [32]. Intercropping with turmeric or Marigold is useful to check the wilting of guava [33].

Biocontrol is the reduction of the amount of inoculum or disease producing activity of a pathogen accomplished by or through one or more organisms other than inoculum. It is the use of natural or modified organisms, genes or gene products to reduce the effects of pests and diseases.

A. Chemical control During 1949, control of wilt was suggested with Chaubatia paste but this control measure is not considered valid, as guava wilt is a soil borne disease. Later 0.1% water-soluble 8-Quinolinol sulphate was found to have chemotherapeutic action against the wilt pathogen (Fusarium oxysporum f. sp. psidii). It’s injection in apparently healthy guava plants in a diseased area provided protection against wilt at least for one year and when injected into slightly wilted plants, it was beneficial for their partial recovery[25]. Many scientists worked on the chemical control of the disease Severe pruning and then drenching with 0.2% either Benlate or Bavistin 4 times in a year and spraying twice with Metasystox and Zinc sulphate were advised but due to soil borne nature of the disease, pruning does not seem to control wilt. It was observed that Benomyl was more strongly absorbed by loamy soil than sandy soil where as in another laboratory test Afugan, Benlate and Topsin were very effective against F. oxysporum f.sp. psidii [26]. Besides, it was reported that among the 10 antibiotics tested against the guava wilt pathogen in in-vitro, actidione was the most toxic, preventing growth even at the lowest concentration of 100fg/ml [27]. Heavy metals Hg, Cd and Cu were also found effective for control of wilt. Different fungicides viz. bavistin, topsin M, indofil M-45, thiram, blitox check the various wilt pathogens in laboratory effectively but these pathogens increases it’s aggressiveness with profuse spore mass production in the soil, once the effect of these fungicides diminishes in soil [28]. The radial growth of Fusarium oxysporum f. sp. psidii was inhibited at high concentrations of carbendazim 50% WP and at low concentration of carbendazim 50%

Aspergillus niger was also found very fast growing, easy to propagate and most effective in controlling the wilt disease in field. Besides this quality, it is also growth enhancer and the plants treated with Aspergillus niger developed faster with more height, more thickness and more numbers of leaves [34]. Out of three bioagents Trichoderma harzianum, T. viride and Gliocladium virens, T. viride is best for the control of wilt [35]. Bioagents like Aspergillus niger, Trichoderma sp. Penicillium citrinum and some bio-dynamic antagonists have shown their effectiveness towards the control of wilt pathogens of guava. Aspergillus niger is found to be most effective in controlling the wilt disease followed by Trichoderma viride [36]. When relative growth of the three bioagents was studied, it was found that Aspergillus niger was fastest growing and most effective [37]. These can be grown easily on any substrate like maize/bajra seeds etc. and can also be multiplied on cheap substrates like Sacchrum sp. (grass) and dry and green leaves of Psidium guajava [38]. The efficacy of Streptosporangium pseudovulgare in controlling rot of guava caused by Lasiodiplodia theobromae was also reported [39]. Co-inoculation of the pathogen with the biological control agent completely inhibited the growth of the pathogen as indicated by the disappearance of the fungal mycelium. The growth of the pathogen immediately stopped following inoculation with the biological control agent. When these fungi were tested 10


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for the control of wilt pathogen in laboratory conditions, these were found quite effective [40]. It was also found that at village level these bioagent can be multiplied in earthen pots [41].

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Management of Fusarium wilt is not possible when we go for a single control measure. So, an integrated approach based on biocontrol agents, plant products and cultural methods are being adopted for the successful management of Fusarium wilt.

Isolates of bioagents, comprising Aspergillus niger, Trichoderma sp. (T. virens, T. harzianum and T. viride) and Penicillium citrinum, their culture filtrates and volatile compounds were evaluated against isolates of F. oxysporum f sp. psidii and F. solani. All isolates of bioagents significantly checked the growth of F. oxysporum f. sp. psidii and F. solani. Per cent inhibition was maximum by direct use of Trichoderma sp. in dual culture A. niger expressed moderate efficacy against F. oxysporum f. sp. psidii and F. solani. However, P. citrinum isolate showed lesser effect. They also observed that in overall, T. virens and T. viride were superior in inhibiting the growth of both Fusarium sp. and Trichoderma and Aspergillus isolates were evaluated under field conditions for the management of guava wilt disease. These bioagents were effective in complete suppression of wilt incidence [42]. The use of VAM symbiont at the rate of 5 kg tree-1 is very beneficial for the control of wilt [43].

A technology has been developed to integrate the different control measures. These includes bioinoculants,viz. Aspergillus niger strain AN 17, Trichoderma harzianum and Penicillium citrinum, intercropping with Curcuma domestica or Tagetes erecta, separate basin irrigation, and avoidance of tillage during July to November and use of resistant root stock. If, these management practices are integrated, the guava wilt could be managed successfully [47]. VIII. DISCUSSION AND CONCLUSION Wilt symptoms start from 28-30 days after inoculation and during September-October fast wilting occurs, while maximum wilting occurs in the month of October. Quantification clearly indicates that October is the most favorable month for wilt incidence which indicates that the increased disease incidence on guava in the subtropics is primarily a function of the guava plant being more vulnerable to infection under rainy/winter temperatures, rather than the pathogen becoming more competitive. In general maximum plants takes three month period for typical wilting after appearance of first visible symptoms though maximum time taken for complete wilting was 240 days.

D. Botanicals An alternative approach in disease management is therefore, the use of botanical fungicides. Plants are the reservoirs of biodegradable secondary metabolites that are reported to inhibit various phytopathogenic fungi. Botanical fungicides developed from plant products are eco-friendly and found to control many plant diseases.

The aqueous extracts/leaf from Curcuma longa L., Achyranthes roses, Calotropis gigantea L. R. Br. Cannabis sativa L. may be more useful against Fusarium wilt pathogens of guava. The extracts/leaf of these plants can be mixed to the soil near root zone of wilted guava plant to control the wilt problem. Meanwhile consortiums of Trichoderma sp. as biocontrol agent guava wilt pathogen viz. F. oxysporum f. sp. psidii and F. solani may be further tested and used for effective management of the disease.

The indiscriminate and inappropriate uses of synthetic fungicides lead to the development of resistance in pathogens, environmental pollution and food contamination by toxic residues. Antifungal activity was reported in leaf extract of Lantana and Azadirachta indica A. Juss. against Fusarium solani, which causes leaf blight of Terminalia catappa [44].. Seventeen plant species were tested including Azadirachta indica A. Juss., Curcuma longa L., Ocimum sanctum along with Datura and Isabgol against wilt of Cumin caused by Fusarium oxysporum f.sp. cumini. and resulted that extract of Datura and Isabgol were more effective fallowed by Azadirachta indica A. Juss., Ocimum sanctum and Curcuma longa L [45]. 23 plant extract were tested in vitro for their biocontrol efficacy to inhibit the growth of Fusarium oxysporum f. sp. psidii and Fusarium solani isolates. Percent of inhibition of fungal growth was calculated as compared to growth in control. Highest percentage of inhibition for all the five isolates of Fusarium oxysporum f. sp. psidii was achieved by extracts from Achyranthes roses, Curcuma longa L. and Calotropis gigantea L. R.Br. and Tagetes erecta L. It was also found that F24 and F30 isolate were comparatively more aggressive among all the isolates of Fusarium oxysporum f. sp. psidii. Leaf extract of Calotropis gigantea L. R.Br and Cannabis sativa L. showed comparatively maximum inhibitory effect to Fusarium solani [46].

Hence, the present investigation illustrates that as the disease is soil-borne in nature, there are a number of limitations for the control of the disease. It is spreading very fast in India and therefore considered to be a serious national problem. Due to high levels of pathogenic and genetic variability the control measures also vary in different parts of India. Further, there is a need for intraand interspecific identification of Fusarium wilt pathogens of guava in order to increase the understanding of the control measures for improving crop productivity. These results can be explored for developing integrated strategies for disease management. IX. REFERENCES [1 ] S.N. Das Gupta and J.N. Rai, Wilt disease of guava (P. guajava). Curr. Sci., 1947, vol.16, 256-258. [2] G. Singh, High density planting in guava- application of canopy architecture. ICAR News (April-June), 2005, vol.11 (2), 9-10.

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[3] V.S. Jaiswal and M. N. Amin, Guava and Jack fruit. In Biotechnology of Perennial fruit crops (End). F.A. Hammerselag and R.E. Litz. CAB international, Wallingford U.,. 1992, pp.421-431.

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[21] R.S. Mathur and S.S. Jain, Selecting guavas for wilt resistance. Proc. Natl. Acad. Sci. B., 1960, vol. 30, 33-36. [22] U.R. Singh, L. Dhar, and G. Singh, Note on performance of guava cultivars and Psidium species against wilt disease under natural field condition. Haryana J. Hort. Sci., 1977, vol. 6, 149-150.

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[33] D.Prasad, S. K. Shukla, B. Prasad and A.K. Misra, Effect of intercrop and different doses of NPK on the incidence of guava wilt. Proceedings of the Seminar on Plant Diseases of National Importance with Special Reference to Guava Wilt and Mango Malformation, April 45, CISH and IISR, Lucknow, 2003, pp. 53-53.

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[17] V.K. Gupta, Molecular characterization of Fusarium wilt pathogens of guava (Psidium guajava L.) using RAPD and microsatellite maker. Ph.D. Thesis, CISH-AU, UP, India. 2010. [18] P.R. Mehta, Observations on new and known diseases of crop plants in Uttar Pradesh. Plant Prot. Bull., 1951, vol. 3, 712-712.

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[19] P.K. Sen and B.S. Verma, Studies on the die-back disease of guava (Psidium guajava L.), A survey of the incidence of the disease in Jhargram area (West Bengal). Proceedings of the 41st Indian Science Congress, (ISC'54), India, 1954, pp. 25.

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[40] G. Neelima, O.M. Prakash, R.K. Pathak, N. Garg and O. Prakash. Biocontrol of rot of guava by Streptosporangium pseudovulgare of cow dung origin. Farm Sci. J., 2003, vol. 12(2), 162.

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[44] A.K. Srivastava, R. Ahmad, S. Kumar and S. Mohandos, Role of VA -Mycorrhiza in the management of wilt disease of guava in the alfivols of Chotanagpur. Indian Phytopathol., 2001, vol. 54, 78-81.

[41] A.K. Misra, D. Prasad, B. Prasad and S.K. Shukla, Effective management of wilt disease of guava. Proceedings of the National. Symposium on Crop Surveillance, Disease Forecasting and Management, Feb. 19-21, Div. of Plant Pathology, IARI New Delhi, 2004, pp. 92-93.

[45] T. Mamatha, and V.R. Rai, Evaluation of fungicides and plant extracts against Fusarium solani leaf blight in Terminalia catappa. J. Mycol. Plant Pathol., 2004, vol. 34, 306-307.

[42] A.K. Misra and D. Prasad, Aspergillus niger strain AN 17 potent bioagent to control wilt disease and its easy multiplication. Proceedings of the Symposium on Recent Advances in Fungal Bioagents and Their Social Benefits, Sept. 10-10, NBRI, Lucknow, 2004, pp. 12.

[46] K. Bhatnagar, B.S. Sharma and H.S. Cheema, Efficacy of plant extract against Fusarium oxysporum f. sp. cumini wilt in cumin. J. Mycol. Plant Pathol., 2004, vol. 34, 360-361.

[43] V.K. Gupta and A.K. Misra, Efficacy of bioagents against Fusarium wilt of guava. J. Mycol. Plant Pathol., 2009, vol. 39, 101-106.

[47] V.K. Gupta, A.K. Misra, B.K. Pandey and U.K. Chauhan, In vitro evaluation of leaf extracts against Fusarium wilt pathogens of guava (Psidium guajava L.). J. Eco-Friendly Agric., 2007, vol. 2, 167-169.

****************** She is currently working as a Research Scholar , Department of Biotechnology , Manav Rachna International University , Faridabad , Haryana , India-121001 M/S Sadana.

Place: Faridabad. Msc in Botany, 2001,University of Rajasthan, Jaipur, Rajasthan, India-302004.

Place: Faridabad. M.tech in Biotechnology, Manav Rachna International University, Faridabad, Haryana , India-121001. Major Field of study: Plant Pathology.

PhD in Plant Pathology, University of Rajasthan, Jaipur, Rajasthan in 2005.

She is currently working as a Research associate, Department of Environmental Biotechnology, Lahti University of Applied Sciences, Finland.

She is currently working as an Assistant Professor, Department of Biotechnology, Faculty of Engineering and Technology, Manav Rachna International University, Faridabad, Haryana, India.

M/S Katyal.

Dr. Didwania Membership: Seminar & Conferences attended (National & International): 25

Place: Faridabad. Msc in Biotechnology , Vinayaka Missions University , Salem, Tamilnadu , India-636308 Major Field of study: Plant Pathology.

13


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