Journal of Scientific & Industrial Research Vol. 76, October 2017, pp. 654-661

Studies on Effect of Grafting Technique on Growth and of Bitter ( Charantia L.)

N A Tamilselvi1* and L Pugalendhi2 *1Department of Vegetable Crops, Horticulture College and Research Institute, Agricultural University, Coimbatore-641 003, Tamil Nadu 2Horticultural Research Station, Yearcaud-636 601 Tamil Nadu

Received 14 August 2016; revised 17 April 2017; accepted 22 August 2017

The experiment was conducted during 2012-14 to study the effect of grafting technique with different rootstocks on growth and yield of bitter gourd ( L.) by comparing grafted bitter gourd with non-grafted (intact) plants under field conditions. The bitter gourd cultivars viz., Palee F1 and CO 1 were grafted onto six cucurbitaceous rootstocks by adopting side grafting method. At 15 days after grafting, the bitter gourd scions grafted with pumpkin (Cucurbita moschata) and gourd (Luffa cylindrica) rootstocks recorded the higher success percentage (89.05 and 85.35) than other graft combinations. The result revealed that, the grafted plants had better vegetative growth and yield than the non-grafted plants (check). Highest vegetative growth (844.26 cm), least node number to first female flower appearance (25.80), narrow sex ratio (17.89) number more fruit number (28.02 fruits) and fruit yield per (3.55 kg/vine) were observed in Palee F1 scion grafted onto ‘pumpkin (Cucurbita moschata)’ rootstock than other graft combinations and non-grafted plants. The differences recorded for growth and yield of the tested cultivars might be due to different growth characteristics, graft affinity and compatibility. This evidence indicates that grafting bitter gourd scions onto specific rootstock significantly influences growth and yield without deterioration in the fruit quality.

Keywords: Grafting, Bitter Gourd, Rootstocks, Yield, Cucurbita moschata, Luffa cylindrica

Introduction Materials and methods Bitter gourd (Momordica charantia L.) is one of Experimental site and growing conditions the important cucurbitaceous vegetable grown in The research was carried out from 2012 to 2014 in . The crop is cultivated over an area of 80990 ha the experimental farm of the Department of in India with an annual production of 830450 tonnes Vegetable Crops, Tamil Nadu Agricultural University, and the productivity of 10.25 t/ hectare1. The main Coimbatore, India at 110 N latitude, 770 E longitudes problem with bitter gourd production in India is 2 and an altitude of 426.26 m above mean sea level. root knot nematode and Fusarium wilt . Several studies on effects of grafting on growth and Rootstocks and scions fruiting characteristics under normal as well as The two bitter gourd cultivars (Palee F1 and CO 1) stress conditions have been conducted in cucurbits. were selected, out of which one is a private hybrid viz., However to our knowledge, there is no published Palee F1 from East West Seeds, which is scientific research on the effects of grafting on suitable for early and long duration harvest and widely increasing resistance to soil-borne diseases as well as cultivated F1 hybrid in Tamil Nadu, India. Fruits are improving yield and yield related characters in bitter medium long with moderate, but rather thick spines gourd in India. Therefore, the hypothesis of this study which result in less damage during transport. CO 1, a was to determine the effects of grafting technique in variety from Tamil Nadu Agricultural University. bitter gourd (Momordica charantia) cv. Palee F1 and Fruits are dark green, 20-25 cm long, susceptible to CO 1 with pumpkin and sponge gourd as rootstocks root knot nematode and Fusarium wilt. The most and ascertain the plant growth, yield and quality. commonly used rootstocks for bitter gourd are Luffa (Luffa spp.), fig leaf gourd (Cucurbita ficifolia) and pumpkin (Cucurbita moschata) showing good —————— 3 *Author for Correspondence vigour and compatibility . In addition pumpkin exhibit E-mail: [email protected] resistant reaction to Fusarium wilt and improves plant TAMILSELVI & PUGALENDHI: STUDIES ON EFFECT OF GRAFTING TECHNIQUE ON GROWTH OF BITTER GOURD 655

vigour; tolerant to squash borer, root knot nematode, The grafting was done under green house during low temperature and drought through vigorous root morning and evening hours. The grafted plants were system. Whereas Luffa is highly compatible, better transferred immediately to the mist chamber where rootstock for earliness, fruit yields, resistant to Fusarium optimum RH (>95per cent), temperature (25-30° C) wilt and tolerance to heat and flooding and widely used and dark condition prevailed for five to seven days. for summer production. The rootstocks Kumatikai After acclimatization, the plants were then transferred (Citrullus colocynthis) and African horned cucumber to shade house on 7th day after grafting. Stock growth (Cucumis met uliferous) found to be highly resistant to and cotyledonary leaves are removed before planting drought, Fusarium wilt and tolerant to root knot in field. Graft compatibility was assessed in terms of nematode. percent survival of grafted plants at 15 days after Fig leaf gourd (Cucurbita ficifolia) and Zucchini grafting (DAG) squash (Cucurbita pepo) were highly tolerant to low temperature and resistant to Fusarium wilt4,5. Based on Preliminary evaluation of graft combinations under field these reports the following rootstocks were selected for condition bitter gourd grafting. The rootstocks viz., Mithipakal About four graft combinations viz., Palee F1 grafted (Momordica charantia var. muricata), teasle gourd onto pumpkin (C. moschata) rootstock, CO 1 grafted (Momordica dioica), athalakai (Momordica cymbalaria), onto pumpkin (C. moschata) rootstock, Palee F1 kumatikai (Citrullus colocynthis), African horned grafted onto sponge gourd (L. cylindrica) rootstock and cucumber (Cucumis met uliferous), fig leaf gourd CO 1 grafted onto sponge gourd (L. cylindrica) (Cucurbita ficifolia), pumpkin (Cucurbita moschata), rootstock recorded maximum success percentage when zucchini squash (Cucurbita pepo), bottle gourd compared to other graft combinations. These graft (Lagenaria siceraria), ash gourd (Benincasa hispida), combinations and thirty days old seedlings of scions ridge gourd () and sponge gourd (Palee F1 and CO 1) were transplanted in the beds at 2 (Luffa cylindrica) were collected from different x 1 m apart under pandal system. The experimental parts of Tamil Nadu, India and grafted with the design consists of randomized block design (RBD) aforementioned two bitter gourd scions. with four replication. There were 25 plants per treatment per replication. Plants were trained over the Screening of rootstocks and scions against meloidogyne pandal after the development of lateral branches and incognita under in vitro Pot culture experiment was conducted under these were vertically supported by jute threads 10 cm glasshouse condition at the Department of Nematology, from the ground level and tied to iron wires distributed TNAU, Coimbatore. Thirty days old healthy seedlings along each row. The shoots arised below the pandal of rootstocks and scions were planted in earthen was eliminated. Recommended package of practices of TNAU was followed to grow a successful crop of pots containing 2.5 kilogramme of sterilized pot mixture 7 (Red soil: Sand: FYM in 2:2:1 ratio).Each pot was bitter gourd . During the growing season, the following inoculated with second stage juvenile of M. incognita vegetative and yield traits were recorded for each plant; vine length (cm), number of primary branches, days to at the rate of two juvenile (J2)/g of soil.The nematode first female flower appearance, node number to female inoculum (J2) was placed in 2 cm depth near to the rhizosphere and covered with sterile sand. The roots of flower appearance, sex ratio, fruit number per vine, these plants were harvested 45 days after inoculation fruit weight (g), fruit length (cm) and girth (cm), flesh and washed free of soil. The observation viz., Number thickness (cm), fruit yield per vine (kg), total number of galls, number of egg masses, number of root knot of harvest and crop duration. The quality parameters nematode females and root knot index were recorded. viz., total soluble solids of fruit samples at vegetable The root knot nematode index (RKI) was calculated by maturity were estimated using hand refractometer and using standard method6. expressed in ºBrix; Ascorbic acid of fruit was estimated by volumetric method and expressed as mg per Grafting 100 g8. Total chlorophyll content of the leaves was Grafting was performed in 25-30 days old seedlings. estimated and expressed as mg per 100 gram9. All the The grafting method adopted in this experiment was statistical analysis was done using AGRES Version side grafting. The best time for side grafting is when 7.01 software. The test of significance was worked out at the rootstock and scion stems have the same diameter. P = 0.05 level of probability10. 656 J SCI IND RES VOL 76 OCTOBER 2017

Results and Discussion cucurbitaceous species against M. incognita (Table 1). All the rootstocks and scions developed The results revealed that kumatikai (C. colocynthis), characteristic galls caused by M.incognita. Significant African horned cucumber (C. metuliferus) and differences were noticed among the rootstocks for pumpkin (C. moschata) rootstocks had the least count number of galls per five gram root, number of egg of root knot index of 2, attaining a reaction category of masses per five gram root and number of females per ‘resistant’ (R), whereas mithipakal (M.charantia var. five gram of root (Table 1). Kumatikai (C. colocynthis) muricata) and sponge gourd (L. cylindrica) exhibited (3.11, 1.42 and 2.34) followed by African horned higher number of galls and recorded a root knot index cucumber (C. metuliferus) (4.20, 1.54 and 2.56) and of 3 and the corresponding reaction category of pumpkin(C. moschata) (8.93, 3.74, 5.56) recorded the ‘moderately resistant’ (MR). The gall index of 4 was lowest number of galls, number of egg masses and observed in ridge gourd (L. acutangula) which attained number of females per five gram of root respectively 11 the reaction category of ‘susceptible’ (S) and other and showed resistant reaction. Siguenza reported rootstocks and scions viz., fig leaf gourd (C. ficifolia), similar results in African horned cucumber zucchini squas (C. pepo), bottle gourd (L. siceraria), (C. metuliferus) and pumpkin (C. moschata). Sponge ash gourd (B. hispida), Palee F and CO 1 recorded the gourd (L. cylindrica) followed by mithipakal 1 root knot index of 5 which is categorized under ‘highly (M. charantia var. muricata) showed moderately resistant reaction and ridge gourd showed susceptible susceptible’. The results of the grafting study revealed reaction. The four rootstocks viz., zucchini squash that seeds of athalakai (M. cymbalaria) and teasle (C. pepo), fig leaf gourd (C. ficifolia), bottle gourd gourd (M. dioica) have not germinated. These are (L. siceraria) and ash gourd (B. hispida) and scions propagated primarily by tubers. The germination and (Palee F1 and CO 1) exhibited highly susceptible growth of kumatikai (C. colocynthis) and African reaction as rhizosphere of these cultivars favoured horned cucumber (C .met uliferus) has found to be too maximum population build up in terms of number of slow. Moreover the stem thickness of these rootstocks galls, number of egg masses and number of females did not match with the thickness of bitter gourd per five gram of root and maximum root damage scions and these were discarded. In case of ash gourd due to gall formation. Similar results were reported (B. hispida) and zucchini squash (C. pepo), quick by Chandra12 in bitter gourd. Root knot index was wilting occurred immediately after grafting. Similar used to evaluate the resistance or susceptibility of incompatibility results were obtained by Miguel 13

Table 1 –– Reaction of wild and cultivated cucurbitaceous species to M. incognita No. of egg No. of RKN Root Knot Degree No. of galls / Rootstock masses/5 g females/5 g Index of resistance 5g of roots of root of root (RKI) Mithipakal 45.80 6.06 30.30 3 MR (Momordica charantia var. muricata) Fig leaf gourd (Cucurbita ficifolia) 129.58 43.20 92.20 5 HS Pumpkin (Cucurbita moschata) 8.93 3.74 5.56 2 R Zucchini squash (Cucurbita pepo) 161.80 50.80 100.90 5 HS Sponge gourd (Luffa cylindrica) 26.36 4.90 12.90 3 MR Ridge gourd (Luffa acutangula) 74.20 25.26 42.23 4 S Bottle gourd (Lagenaria siceraria) 86.40 35.60 46.70 5 HS Ash gourd (Benincasa hispida) 98.40 39.06 55.10 5 HS Kumatikai (Citrulus colocynthis) 3.11 1.42 2.34 2 R African horned cucumber 4.20 1.54 2.56 2 R (Cucumis metuliferus) Scions

Palee F1 76.60 22.66 33.93 CO 1 78.30 26.76 38.20 5 HS SEd 1.81 1.02 1.09 5 HS CD (P=0.05) 3.75 2.10 2.26 *Inoculation level 2J2/gramme of soi TAMILSELVI & PUGALENDHI: STUDIES ON EFFECT OF GRAFTING TECHNIQUE ON GROWTH OF BITTER GOURD 657

when grafting melon onto C. melo var. saccharinus which has direct impact on fruit yield, since plant and Morra14 reported that melon scions showed low population and fruit yield are closely related. About affinity with L. siceraria, C. ficifolia and C. moschata four graft combinations viz., Palee F1 grafted onto rootstock. In the incompatible combinations, the pumpkin (Cucurbita moschata), CO 1 grafted onto leaves turned yellow and plants get withered due to pumpkin (Cucurbita moschata), Palee F1 grafted onto poor vascular connection with poor connecting sponge gourd (Luffa cylindrica) and CO 1 grafted onto sieve tubes, cambium and in the heterograft sponge gourd (Luffa cylindrica) recorded maximum Cucumis/Cucurbita15. Graft success per cent was success percentage when compared to other graft assessed onto six cucurbitaceous rootstocks employing combinations. These grafts were planted in the main th two scions viz., Palee F1 and CO 1 at 15 days after field along with their scions viz., Palee F1 and CO 1 and grafting (DAG) showed significant differences among evaluated for growth, yield and quality parameters. the rootstocks (Figure 1). Among the rootstocks, Growth and flowering behaviour of bitter gourd grafts pumpkin recorded the highest success per cent of 89.0 and scions were presented in (Table 2). The results of followed by sponge gourd which recorded 85.3 per this study revealed that vine length was significantly cent. The lowest graft success per cent (50.5 per cent) higher in grafted plants than that of non grafted was observed in mithipakal rootstock followed by plants. Among the four graft combinations, Palee F1 fig leaf gourd rootstock (45.0 per cent). Successful grafted on to pumpkin (Cucurbita moschata) rootstock grafting (compatibility between scion and rootstock) recorded highest vine length of 844.26 cm at the end may be due to cell division in the scion and of the experiment (Table 2). Similarly vigorous plant rootstock at the graft union site (dedifferentiation and growth was observed in grafted watermelon plants19. redifferentiation of the callus tissue), followed by rapid Promoted plant growth in grafted plants can be connection between the vascular bundles of the scion explained by the interaction of some or all of the and rootstock and subsequent secondary growth of the following phenomena: increased water and plant scion 16. The rootstocks viz., Luffa (Luffa sp.), fig leaf nutrient uptake, due to stronger and more extensive gourd and pumpkin are common rootstocks for bitter root growth of the rootstock. About 32 per cent higher gourd grafting and also noticed that bitter gourd scions main vine length were observed in grafted cucumber grafted onto Luffa recorded higher plant survival per plant than that of non-grafted counterpart20. The cent (87.0 and 81.5 per cent ) than bitter gourd grafted number of primary branches per plant is yet another onto bottle gourd rootstock3,17,18. Contrary to the yield contributing trait in bitter gourd which differed 3 present study Lin reported that cleft grafting is a significantly among the treatments. Palee F1 grafted suitable grafting technique for bitter gourd grafting. onto pumpkin (Cucurbita moschata) rootstock Success percent of graft is an important attribute which recorded the highest number of primary branches influence the plant population under field condition, (12.97) followed by Palee F1 grafted onto sponge gourd (Luffa cylindrica) rootstock (11.01) than the other grafted and non-grafted plants (Table 2). Similarly grafted watermelon plants produced more lateral (9 lateral vines / vine) than non-grafted plants (4 lateral vines/vine)21. Earliness is one of the main attributes which is measured in terms of days to female flower appearance and is preferred for commercial cultivation when high yield is coupled with earliness. The results of this study indicated that grafted plants showed seven to ten days delay in flowering (both male and female) than non-graftedplants due to heavy stress during

graft union process. But Palee F1 grafted onto pumpkin Fig.1 — Effect of rootstocks on grafting success of bitter gourd (Cucurbita moschata) rootstock exhibited earliness scions at 15 days after grafting. Note: T1- Palee F1 /Mithipakal, with less number of days to first female flowering T2 – CO 1 / Mithipakal, T3- Palee F1 / Fig leaf gourd, T4- Palee (70.32) when compared to other graft combinations F1 / Fig leaf gourd, T5 – Palee F1 / Pumpkin, T6 – CO 1 / Pumpkin, (Table 2). Similar trend of delayed flowering up to T7 – Palee F1 /Sponge gourd, T8 –CO 1/Sponge gourd, T9 – Palee F1 /Ridge gourd, T10 – CO 1 / Ridge gourd, T11 – Palee F1 / Bottle one week were reported in grafted watermelon plants 22 gourd & T12 - CO 1 / Bottle gourd. resulting in an equal delay in fruit maturity . 658 J SCI IND RES VOL 76 OCTOBER 2017

Table 2 — Growth and flowering behaviour of grafted and non grafted bitter gourd plants Bitter gourd cultivars and Vine length at Number of Days to Node no. to first Sex ratio Days to graft combinations final harvest primary first female female flower (No. of male/ first harvest Scion / Rootstock (cm) branches flower appearances female flower) appearances

Palee F1 bitter gourd T1-Palee F1 ungrafted 647.67 11.02 63.30 30.12 19.02 77.62 (intact) T2-Palee F1 / Pumpkin 844.26 12.97 70.32 25.80 17.89 83.47 (Cucurbita moschata) T3-Palee F1 / Sponge gourd 511.97 11.01 76.97 33.17 20.67 91.10 (Luffa cylindrica) CO 1 bitter gourd T4-CO 1 non grafted (intact) 543.76 10.27 68.47 34.02 23.09 84.52 T5-CO 1 / Pumpkin 619.42 10.30 81.70 31.21 23.05 96.90 (Cucurbita moschata) T6- CO 1 / Sponge gourd 393.20 9.74 87.20 34.12 28.97 103.32 (Luffa cylindrica) SEd 6.12 0.70 1.12 1.31 1.25 1.02 CD (P=0.05) 13.05 1.50 2.39 2.79 2.67 2.18

Node at which first female flower appearance is also in grafted plants was mainly due to delayed flowering considered as important biometric trait to measure the because of heavy stress during graft union process. The earliness in cucurbits. The graft Palee F1 grafted onto fruit yield and yield attributes varied significantly pumpkin (Cucurbita moschata) rootstock produced among the grafted and non-grafted (scion) plants female flowers at the early nodes (25.80) than the (Table 3). The graft Palee F1 grafted onto pumpkin other grafts and scions (Table 2). This was line with the (Cucurbita moschata) rootstock recorded highest fruit findings of Reyes17 who reported grafting bitter gourd number (28.02) and yield per vine (3.55 kg) which is scions onto sponge gourd and bottle gourd rootstocks significantly differed from the non-grafted plants or had the first female flower at the earlier nodes than that scions. In the present study, fruit number and yield per of non-grafted plants (Table 2). Narrow sex ratio is vine of Palee F1 grafted onto pumpkin (Cucurbita favorable trait in cucurbits. Grafted bitter gourd plants moschata) rootstock were increased by 32 per cent tend to give higher female and lower male flowers and 22 per cent than that of Palee F1 scion (Table 3, when compared to non-graftedplants. In the present Figure 2 & 3). This might be resulted from different investigation, the graft combinations, Palee F1 grafted factor such as vigorous root system of the pumpkin onto pumpkin (Cucurbita moschata) rootstock had rootstocks increases the efficiency of water and the narrow sex ratio (17.89) than that of other grafted nutrient consumption resulting in enhanced growth and and non-grafted plants (Table 2). Similar findings were yield. Superior interaction between rootstock and scion observed in grafted watermelon23. Observation on days on vigour of the root system which could enables good to first harvest is an indicator of the earliness of any supply of endogenous hormones leading to increased crop especially in vegetables which could fetch fruit yield. Similar trends of increased fruit number and premium price and catch the early market. It is directly yield in grafted plants were also reported in grafted influenced by earliness in flowering. Earliness in watermelon19,23. Fruit physical features (fruit weight, flowering is directly proportional to number of length, girth and flesh thickness) recorded in the harvests. Non-grafted plants or scions recorded lowest grafted and non-grafted bitter gourd plants indicated value for days to first harvest than grafted plants. that fruits harvested from grafted plants differed Among the four graft combinations, Palee F1 grafted significantly from those of non-grafted (intact) plants onto pumpkin (Cucurbita moschata) rootstock showed Table 3. The highest mean fruit weight (182.20 g), earliness in the first harvest (83.47 days) which length (25.30 cm), girth (16.25 cm) and flesh thickness recorded high fruit yield (Table 2). Delayed harvesting (0.85 cm) were obtained in Palee F1 grafted onto TAMILSELVI & PUGALENDHI: STUDIES ON EFFECT OF GRAFTING TECHNIQUE ON GROWTH OF BITTER GOURD 659

Table 3 — Yield and quality of grafted and non grafted bitter gourd plants

Bitter gourd ) cultivars and graft -1 TSS combinations (cm) (cm) (cm) (º Brix) Brix) (º per vine per (mg g plant (kg) plant of harvest of Fruit girth Fruit Scion / Rootstock Individual (mg/100 g) (mg/100 Fruit length length Fruit Fruit number number Fruit Total number Total Ascorbic acid Ascorbic Crop duration Crop Fruit yield yield Fruit per fruit weight (g) fruit Flesh thickness Flesh Total chlorophyll chlorophyll Total

Palee F1 bitter gourd T1-Palee F1 ungrafted (intact) 21.22 164.55 20.20 14.97 0.73 2.90 13.17 166.15 104.63 5.10 2.12

T2-Palee F1 /Pumpkin 28.02 182.20 25.30 16.25 0.85 3.55 17.08 194.12 105.87 5.05 2.16 (Cucurbita moschata) T3-Palee F1 / Sponge gourd 19.40 137.32 19.90 15.37 0.75 2.27 14.40 182.75 101.80 4.63 1.89 (Luffa cylindrica) CO 1 bitter gourd T4-CO 1 18.07 92.14 23.17 13.65 0.68 1.79 9.49 128.90 96.80 4.49 1.69 nongrafted (intact) T5-CO 1 / Pumpkin 20.93 134.22 22.87 14.65 0.70 2.20 13.05 154.97 97.02 3.54 1.60 (Cucurbita moschata) T6-CO 1 / Sponge gourd 18.80 101.82 19.32 14.87 0.66 1.70 8.30 144.75 88.63 3.36 1.18 (Luffa cylindrica) SEd 0.71 2.06 1.45 1.11 0.03 0.23 1.15 2.03 1.06 0.13 0.10

CD (P=0.05) 1.53 4.40 3.10 2.37 0.06 0.50 2.45 4.33 2.27 0.29 0.22

pumpkin (Cucurbita moschata) rootstock followed by respect to total soluble solids (TSS) and ascorbic acid Palee F1 grafted onto sponge gourd (19.90 cm, 15.37 content (Table 3). But the differences were not cm and 0.75 cm respectively) except for fruit statistically different between the grafted and non- weight.The fruit weight recorded by this graft grafted (intact) plants. Significant differences were combination was 33 per cent higher than the other observed among the graft combinations with respect to grafted plants and scions. Similar differences in total chlorophyll contents of leaf. The highest total fruit physical features were recorded in chlorophyll content was recorded by Palee F1 grafted watermelon19,23. More number of harvest and long onto ‘pumpkin (Cucurbita moschata) rootstock’. crop duration would give higher fruit yield. In general Significant differences in total chlorophyll content the grafted plants had longer crop duration and suggest the possible role of the scion/rootstock give more number of harvests than that of non-grafted interaction in the noticeable increase detected in these plants. In this study, the graft combination, Palee substances. The increase in the photosynthetic rate F1 grafted onto pumpkin (Cucurbita moschata) might be due to enhanced synthesis of endogenous rootstock recorded significant values for the same traits hormone which attributed to enhanced ultra structural (Table 3). Similar results were noticed in brinjal who morphogenesis of plastids and increased the Rubisco 25 stated that crop duration was one month longer which activity . The overall study indicated that Palee F1 leads to more number of harvests per plant than grafted onto pumpkin (Cucurbita moschata) rootstock that of non-grafted plants24. The physicochemical is potential graft combination to be of valuable characteristic of the fruits showed some variation alternative to get higher yield and profit compared to among the grafted and non-grafted (intact) plants with other graft combinations and scions. 660 J SCI IND RES VOL 76 OCTOBER 2017

Fig.2 — Effect of grafting on fruit number per vine

Fig.3 — Effect of grafting on fruit yield per vine (kg)

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