20017-Amit Kumar 24 June, 2020

Indian Journal of Entomology Online published (Preview) DoI No.:

BIOCHEMICAL BASIS OF RESISTANCE TO MUSTARD ERYSIMI (KALTENBACH)

Amit Kumar*, Sunita Yadav**, Nisha Ahlawat and Jayant Yadav

Department of Genetics and Plant Breeding; *Department of Entomology, CCS Haryana Agricultural University, Hisar **Email: [email protected] (corresponding author)

ABSTRACT

This study evaluates the biochemical basis of resistance to mustard aphid (Kaltenbach) in Brassica genotypes under semiarid conditions of Haryana. The study was carried out during rabi s2016-17 at the research area and laboratory of Oilseeds Section, Department of Genetics Plant Breeding, CCS Haryana Agricultural University, Hisar. Ten genotypes belonging to Brassica spp., viz., B. campestris (BSH-1), B. carinata (HC-212), B. juncea (Purple Mutant), B. tournefortii (DRMR-2205), B. napus (HNS- 9605), B. nigra (Nigra), B. rapa L. var. toria (Sangam), B. rapa L. var. yellow (YSH-401), E. sativa (T-27) and Sinapsis alba (DRMR-2183) were included. Observations on the L. erysimi at flower initiation, full flowering and full pod formation/setting stages and aphid infestation index (AII) was computed. These data were correlated with biochemical parameters such as total phenols, sugars, proline content and leaf waxes. The genotypes viz., BSH-1, YSH-401 and HC-212 were observed with very low phenol content (1.51, 1.60 and 1.85%, respectively). The AII and total phenols were observed significantly negatively correlated (r = -0.92**). The effect of waxes in leaves had a lethal effect (AII vs leaf waxes correlation being significantly negative with r = -0.91**). AII was observed significantly positively correlated with proline content (r =0.91**) and total sugar content (r= 0.98**). Regression analysis revealed that the biochemical contents collectively accounted for 97% variability in aphid infestation.

Key words: Brassica genotypes, Lipaphis erysimi, resistance, biochemicals, phenol, leaf waxes, sugar, proline, correlation coefficients, aphid infestation index

Rapeseed-mustard belonging to Brassica and 31% seed weight loss, and 5-6% reduction in oil (Cruciferae) is one of the important oilseed crops. As a content (Bakhetia and Sekhon, 1989; Singh and Sharma, group, these comprise the traditionally grown indigenous 2002; Dhaliwal et al., 2004; Rana 2005; Shylesha et al., species, namely toria (Brassica rapa L. var. toria), 2006; Parmar et al., 2007). Such losses may go up to brown sarson (B.rapa L. var. brown sarson), yellow 100% in certain mustard growing regions (Singh and sarson (B. rapa L. var. yellow sarson), Indian mustard Sachan, 1999) due to infestation (Mandal et al., 2012). (B. juncea L.), black mustard (B. nigra Koch), taramira Insecticides are mostly used against these , but (Eruca sativa/ vesicaria Mill.) and non-traditional these are harmful to their natural enemies (Singh et species like gobhi sarson (B. napus L.) and Ethiopian al., 2007), and also cause pollution, residues and other mustard or Karan rai (B. carinata Braun). Haryana is hazards (Singh and Sharma, 2002). Use of resistant the third most important rapeseed-mustard producing cultivars is an ecofriendly alternative IPM strategy as state with a productivity of 1721 kg/ha (2015-2016) it is compatible with other control methods. This study (Agricultural statistics at a glance, 2017). Among the evaluates the effect of biochemicals in Brassica spp., constraints in productivity, the damage done by on aphid resistance. pests is most important. PreviewRai (1976) listed 24 species, while Bakhetia and Sekhon (1984) enlisted 38 species. MATERIALS AND METHODS However, Purwar et al. (2004) reported more than 43 The field experiment was carried out at the research species of insect-pests out of which about a dozen area and laboratory of Oilseeds Section, Department of are considered as major pests. Among these aphids Genetics Plant Breeding, CCS Haryana Agricultural [Lipaphis erysimi Kalt., L. and University, Hisar (29010’N, 75046’E, 215 masl) Sulzer (: )] are the during rabi 2016-17. The crops were grown under late most destructive (Desh Raj et al., 1996; Sarangdevot et sown (date of sowing 27.11.2016) conditions with al., 2006). Of these, L. erysimi causes up to 9-96% yield Recommended Package of Practices (Anonymous, 2 Indian Journal of Entomology Online published (Preview)

2015). Ten genotypes belonging to ten Brassica spp., Plant biochemical constituents may have role in in plots of 2x 3 m, with spacing of 30x 10 cm, in RBD imparting resistance to the plants against mustard aphid. with three replications. The Brassica spp. viz., B. So five randomly selected competitive plants of each campestris (BSH-1), B. carinata (HC-212), B. juncea genotype were uprooted at bud + inflorescence stage (Purple Mutant), B. tournefortii (DRMR-2205), B. from the field and separated into leaves and bud + napus (HNS-9605), B. nigra (Nigra), B. rapa L. var. inflorescence portions. The samples were brought to the toria (Sangam), B. rapa L. var. yellow (YSH-401), laboratory, kept in airtight plastic containers, and stored E. sativa (T-27) and Sinapsis alba (DRMR-2183) at 40C. Total phenols, total sugars, proline content and were collected from different centers of the All India leaf waxes were estimated by the methods as suggested Coordinated Research Project (AICRP) on Rapeseed by Swain and Hills (1959), Yemm and Willis (1954) and Mustard, Sewar, Bharatpur (Rajasthan). using anthrone reagent, Bates et al. (1973) and Ebercon et al. (1977) by colorimetric analysis, respectively. Aphid Infestation Index (AII) was computed with Estimation of total phenols, total sugars and proline the observations made on the population of L. erysimi at content was done for both leaves and inflorescence flower initiation, full flowering and full pod formation/ parts but wax content was estimated only for the leaves setting stages. Ten plants were selected at random from of plants because waxes are the main constituents of each in each replication, and number of aphids observed leaves and their quantity is negligible in inflorescence. from top 10 cm portion of the terminal shoot (Bakhetia Correlation and multiple regression analysis of AII and Sandhu, 1973). Finally, the AII of the three stages with biochemical parameters was done with Online were pooled and the pooled mean was calculated. For Statistical Analysis Package (OPSTAT) by Sheoran et AII, the numbers of selected/ tagged plants falling in al. (1998). Also, the AII was plotted in the graphs along each grade were multiplied by respective grade and the with the biochemical constituents. total was divided by plants population in each repeat of a genotype on which grading was done. The pooled RESULTS AND DISCUSSION mean I of each genotype was workout on the basis of the mean of three replications. Biochemical constituents (Table 1) The amount of phenol content in leaves varied (0 ´ a) + (1 ´ b) + (2 ´ c) + (3 ´ d) + (4 ´ e) + (5 ´ f) ______from the minimum of 1.25% in BSH-1 to maximum All = a + b + c + d + e + f of 3.04% in Nigra. Similar trend was observed in the inflorescence of various Brassica genotypes with Here, a, b, c, d, e and f are number of plants the minimum of 1.77% phenol content in BSH-1 to (frequency) falling in each grade. The grades as maximum of 3.33% in genotype Nigra. On the basis of described by Bakhetia and Sandhu (1973) were pooled mean of plant parts, total phenols varied from distributed as: the minimum of 1.51% (BSH-1) to maximum 3.19% (Nigra). The genotypes such as T-27, Sangam and Nigra Grade Description had phenol content more than the mean i.e. 2.29%. 0 Free from aphid infestation. Plants show excellent The genotypes viz., BSH-1, YSH 401 and HC 212 had growth. 1 Plants having 1-15 aphids/ inflorescence. Normal very low phenol content on the basis of pooled mean growth, no curling or yellowing of a few leaves, of plant parts i.e. 1.51, 1.60 and 1.85%, respectively. except only a few aphids along with little or no Leaf waxes (%): The amount of wax content in leaves symptoms of injury. of Brassica genotypes varied from the minimum of 2 Plants having 16-100 aphids/ inflorescence. Average 2.89% (BSH-1) to the maximum of 5.72% (T-27). Table growth, curling and yellowing of a few leaves. 1 showed that the genotypes having high leaf waxes 3 Plants having morePreview than 100 aphids/ inflorescence. viz., Purple Mutant (4.92%), DRMR-2205 (5.35%), Growth below average, curling and yellowing of the leaves on some branches. Plants show some Sangam (5.39%), Nigra (5.45 %) and T-27 (5.72 %) had stunting, poor flowering and little pod setting. low infestation of mustard aphid (0.8-1.6 AAII). The 4 Heavy aphid colonies on plants. Very poor growth, genotypes such as BSH-1, HC 212, YSH 401 and HNS heavy curling and the yellowing of leaves, stunting 9605 had lowest leaf wax content i.e. 2.89, 3.1, 3.17 and of plants, little or no flowering and only a few pods 3.21%, respectively. Proline (µmol/g FW) content in forming. leaves of Brassica genotypes varied from the minimum 5 Plants full of aphids. Heavy stunting of plants; of 15.39 (Nigra) to the maximum of 18.25 µmol/g curling, crinkling and yellowing of almost all the leaves. No flowering and pod formation. (BSH-1) whereas in the inflorescence, it varied from Biochemical basis of resistance to mustard aphid Lipaphis erysimi 3 Amit Kumar et al. 7.64 9.58 7.69 8.71 8.77 8.85 8.08 8.87 7.71 7.64- 10.73 10.76 10.76 (%) (0.53) Total Total 0.98** sugars

13.38 16.36 15.80 15.25 13.02 14.98 14.67 14.55 14.28 16.36 15.30 12.42 (0.06) 12.42- FW) 0.91** Proline (µ mol/g Pooled data 2.86 1.51 1.60 1.85 2.85 2.03 2.21 2.29 2.71 3.19 2.07 3.19 1.51- (%) (0.17) Phenols Y= -15.20 + 0.94 TP + 0.44 TP Y= -15.20 + 0.94 TS PRO + 0.99 0.97 -0.92** 8.12 8.16 9.23 9.30 9.34 8.29 9.41 8.24 8.12- 11.24 11.43 11.43 10.01 (0.42) (%) 0.97** Total Total sugars Brassica genotypes 9.45 9.45- 10.81 14.46 14.21 13.93 10.50 13.16 13.35 12.54 10.96 14.53 14.53 (0.05) FW) 0.84** Proline (µ mol/g Flower + flower bud 2.95 1.77 1.85 2.04 3.05 2.08 2.25 2.41 2.66 3.33 2.13 3.33 1.77- (%) (0.19) -0.88** Phenols Y= -4.64 - 0.31 TP - 0.01 PRO TP Y= -4.64 - 0.31 + 0.81 TS 0.94 7.15 9.15 7.22 8.18 8.24 8.36 7.86 8.32 7.18 7.15- 10.22 10.09 10.22 (0.63) (%) Total Total 0.98** sugars

18.25 17.39 16.57 15.54 16.80 15.99 16.55 17.59 18.25 16.07 0.75* 15.39 15.95 (0.07) 15.39- FW) Proline (µ mol/g Leaves 2.89 3.17 3.10 5.72 4.92 4.26 4.35 5.35 5.72 3.21 5.45 5.39 2.89- (%) (0.29) Leaf waxes -0.91** 1.25 1.34 1.66 2.64 1.97 2.16 2.16 2.76 3.04 2.01 3.04 2.76 1.25- (%) (0.14) -0.94** Phenols Phenols Y= -5.19 + 0.12 TP - 0.26 WX + 0.09 - 0.26 TP Y= -5.19 + 0.12 TS PRO + 0.80 0.98 — — — — 4.3 3.6 2.9 0.9 1.6 2.2 1.5 2.2 0.8 1.1 AII Table 1. Biochemical constituents in relation to mustard aphid resistance of Table Genotypes BSH-1 YSH-401 HC-212 T-27 Purple Mutant DRMR-2183 — DRMR-2205 — — HNS-9605 — Nigra PreviewSangam Brassica spp. B. campestris . var. B. rapa L . var. yellow B. carinata Braun . Eruca sativa Mill B. juncea Sinapsis alba Mean B. tounefortii Range CD at (p=0.05) B. napus ‘r’ Regression equation R2 B. nigra Brassica rapa toria L . var. 1. 8. 2. 9. 3. 4. 5. 6. 7. S. 10. No. Figures in parentheses angular transformed values, ** significance at p = 0.01, *significant at p Aphid= 0.05; Y= Infestation Total Index, Phenols, Waxes, TP= WX= PRO= Leaf Sugars Total TS= Proline Content and 4 Indian Journal of Entomology Online published (Preview) the minimum of 9.45 µmol/g (Nigra) to maximum of and glucosionates. Similar results were obtained by 14.53 µmol/g (HNS 9605). On the basis of pooled mean Kular et al. (2008) that the concentration of phenols of plant parts, proline content varied from the minimum was lowest in purple mutant as compared to T-27. of 12.42 µmol/g (Nigra) to maximum 16.36 µmol/g Contrary to the observations of Narang (1982), Gill and (BSH-1). On the basis of pooled mean it is evident Bakhetia (1985) observed a negative and non-significant that the genotypes such as DRMR 2183, Purple Mutant correlation of phenol content with aphid incidence. The and HC 212 had proline content more than the mean effect of waxes in leaves had lethal effect, and similar i.e. 14.55 µmol/g. The genotypes such as Nigra, T-27, results were obtained by Yadav et al. (1985). With B. Sangam and DRMR 2205 had lowest proline content campestris (BSH-1) and B. juncea (Parkash) heavy i.e. 12.42, 13.02, 13.38 and 14.28 µmol/g, respectively. infestation was observed and these were categorized as susceptible. Present results are in line with that of Total sugars (%): The amount of total sugars varied Ahlawat et al. (2008), and Kumar (2008) as regards wax from 7.15% (Sangam) to 10.22% (BSH-1) in leaves content in leaves varying from 2.91% (BSH-1) to 5.72% (Table 1) and in inflorescence it varied from 8.12% (T-27) and concluding that waxes content in leaves is (Sangam) to 11.43% (YSH 401) (Table 1). On the negatively but non-significantly correlated. However, basis of pooled mean total sugar content varied from these results contradict with those of Kundu and Pant 7.64% (Sangam) to 10.76% (YSH 401) (Table 1). The (1967), who did not observe any correlation between genotypes viz., BSH-1, YSH-1 and HC 212 had highest developmental period of L. erysimi with nitrogen, total amount of total sugars in leaves i.e. 10.22, 10.09 and sugars, amino acids and amides content. 9.15% respectively whereas in inflorescence stage Brassica genotypes viz., YSH-401, BSH-1, HC-212, Cumulative effect of biochemicals on the AII, when HNS-9605 and DRMR-2183 had highest amount of analysed with regression coefficients, it was observed total sugars i.e. 11.43, 11.24, 10.01, 9.41 and 9.30%, that none of the biochemical traits is solely responsible respectively. for aphid infestation (Table 1); all the biochemicals collectively accounted for 98 and 94% variability Correlation of Aphid Infestation Index in infestation in case of leaves and inflorescence, The correlation analysis between various biochemical respectively. Pooled data showed that biochemicals in traits present in leaves and AII as shown in the Table 1 both leaves and inflorescence collectively accounted revealed that phenols and leaf waxes were negatively for 97% variability. and significantly correlated (r = -0.94** and r = -0.91**) with AII. The correlation of AII was also significant and REFERENCES positive with proline content (r = 0.75*) and total sugars Agricultural Statistics at a Glance. 2017. Government of India, Ministry (r =0.98**). Similarly the correlation analysis between of Agriculture, Department of Agriculture & Cooperation, various biochemical traits in flower + flower bud and Directorate of Economics & Statistics, Krishi Bhawan, New Delhi. AII presented in the Table 1 revealed that AII and Ahlawat D S, Singh H, Singh D, Rohilla H R. 2008. 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(Manuscript Received: ______, 2020; Revised: ______, 2020; Accepted: ______, 2020; Online Published: ______, 2020) OnlinePreview published (Preview) in www.entosocindia.org Ref. No. 20017