Response of Dragon Fruit (Hylocereus sp.) Cuttings to Different Plant Growth Regulators.

Introduction

Dragon fruit (Hylocereus sp.) has received worldwide recognition ¹ as a promising fruit crop.  It belongs to family Cacteceae which comprises of 120-200 genera and 1500-2000 species with  their existence in  America². Due to the formation of large flowers blooming at night dragon has an ornamental value. The fruit comes in the market in three forms with leathery, slightly leafy skin. Due to the presence of bracts or scales on fruit skin the fruit has been given the name of pitaya. It is also known as pitahaya, pitajuia, pitalla or pithaya³.The fruit is mainly confined to tropical and subtropical regions of Mexico and Central south America⁴. Besides industrial as well as  medicinal significance it also excels economic potential due to which this fruit crop has been taken up for commercial cultivation by the grower’s world-wide⁵. Dragon fruit is packed with potential health benefits. Recent studies investigate the antioxidants activity of dragon fruit which suggested the pigments in providing protection against certain oxidative-stress related disorders⁶. A broad spectrum of vitamins, lycopene, minerals and carbohydrates in the form of reducing sugars including glucose and fructose has been advocated^7,8. The pulp and peel extract showed anti-inflammatory, anti-spasmodic, radioprotective⁹ antimicrobial, anti-cancer and anti-diabetic activities¹⁰. Both the flesh and seeds have a good control on fatty acids. Dragon fruit is eaten as a fresh fruit. Fresh and dried dragon skin is used as natural food thickener and natural coloring agent. Albedo of the fruit is used to color rice, milk, yoghurt, juice and pastry¹¹. Propagation of dragon fruit through vegetative method is a pre-requisite to produce true to type plants. Among vegetative propagation cuttings of majority of plants are normally dipped in growth regulations before planting to boost root formation. Among propagation techniques the easiest and cheapest method of propagation is by cutting^{12, 4}. Propagation through cutting ensures faster fruiting. In some dragon fruit cultivars cuttings do not root easily without the use of auxin treatments and have extended root initiation and root growth as compared to treated cuttings. Cuttings of majority plants are normally dipped in growth regulators before planting to boost root formation. Researchers demonstrated the rooting stimulated by IBA treatment^{13, 14}.

Materials used and methodology

Procurement of cuttings and treatment

The dragon fruit cuttings were sourced out of a one year old healthy mother plant of cv. American Beauty. The solution of growth regulators were prepared by dissolving it in 50 % ethanol (with a purity of 96%) with different concentrations. Sixteen treatment combinations (100-500)  ppm NAA, (1000-5000) ppm IBA and (500-1500 ) ppm PHB along with untreated cuttings were replicated three times in randomized block design. The cuttings of the size of 20 cm were planted in the polybags of size of 16×10 inches filled with soil in the middle of March.

Rooting media

Before planting, the polybags of size of 16×10 inches were filled with the respective rooting media. On the bottom and sides of polythene bags, holes for water drainage were made.

Cutting plantation

While planting, about 2/3^rd length of the cuttings were buried in the media, leaving 1/3^rd part exposed in the environment.

Experimental procedures

Days of sprout initiation

Daily observations were  recorded of the treated cuttings regarding sprout initiation and the time elapsed in it was noted .

Number of sprouts

The number of sprouts per cutting was calculated out of the total cuttings planted.

Percentage of shoot emergence

Shoot emergence percentage was calculated by dividing the total sprouted cuttings by total planted cuttings and expressed as its percentage.

Sprout length

Shoot length was measured by a measuring scale taking into account the base up to the growing tip.

Fresh shoot  weight

It was taken on average basis of the randomly selected five plants per treatment on an electronic balance expressed in grams.

Dry shoot weight

Using destructive method the selected shoots were packed in paper bags and  then air  dried  at 60° C in hot air oven.  On  drying, their  weight was calculated as grams.

Survival

The survived cuttings under each treatment in each replication was recorded and its percentage was calculated by the division of total survived cuttings by total planted ones expressing it in per cent.

Roots per cutting

The roots arising from the base of the cuttings were counted after  gentle washing them with water.

Average root length

Starting from the base to growing tip of root length was measured with scale of the selected plants and later on averaged.

Longest root length

Length of longest root was taken as  centimetres after its measurement with the scale.

Fresh root weight

After separation of the roots they were washed and weighed on an  electronic balance and  its average was calculated in grams.

Root  dry weight

To calculate the dry weight of roots, roots were air dried at 55⁰C in hot air oven. After drying the weight was calculated again on the electronic balance and weight was calculated in grams.

Data collection and statistical analysis

The observations pertaining to germination, vegetative growth and survival were calculated at 90 DAP and processed in MS-Excel. The statistical analysis of data which comprised of 7 treatments and 3 replications were analyzed by randomized block design with factorial arrangement (p≤0.05) with  IBM-SPSS Statistics (Version 29.0) software. The significant difference between means were compared by using DMRT’S.

Experimental results and discussion

Number of sprouts

It is apparent from the results that slight variations were found among the treatments on sprout production in the cuttings. Increased sprout production was achieved up to certain concentrations but decreased at the highest concentration (Table-1). Maximum sprouts (2.94) were formed in IBA 4000 ppm. PHB succeeded IBA within the range of 0.99 -1.86 with the maximum (1.86) in PHB 750 ppm which also showed decreased sprout production with increased concentrations . The untreated cuttings produced the least (0.83) sprout production. The accelerated sprout formation can be ascribed to the enhanced division of cells and their elongation with application of increased dose of  IBA leading to an increased shoot growth activation that led to sprout development¹³. The research study of ¹⁶ stated that sprouting can be due to the availability of the reserved food material as recorded in Citrus limon²⁰, in pomegranate ²¹ and in dragon fruit cuttings^{22, 23}.

Percentage of shoot emergence

Based on the data analysis presented in Table-1 the treatments administering NAA, IBA and PHB significantly affected the percentage of shoot emergence. More prominence in shoot emergence was observed in IBA treatments than NAA and PHB treatments. The application of IBA has a significant effect on the percentage of shoot emergence namely (4000 ppm) as the maximum percentage of shoot emergence. In PHB treatments emergence percentage was in the range of 39.78 to 42.09 per cent with an increase in emergence with increasing concentration depicting maximum 51.41 per cent in 1000 ppm treated cuttings. Out of all NAA registered the lowest emergence within the range 53.31-67.33 per cent with an increase up to 400 ppm NAA as maximum (67.33 %) shoots. For promotion  of  adventitious rooting IBA treatment proved to be the most effective²⁴. The increased concentration of IBA increased the root formation²⁵ due to the presence of stable chemical content for root stimulation²³ in dragon fruit cuttings and ensured improved uniformity in plants. The  acceleration of cell activity  resulted from the energy obtained from the proper utilization of the reserved food material might have resulted in the  increased  sprout emergence percentage^{26, 27}. The research findings concluding IBA as the best for the establishment of the dragon fruit cuttings are akin to the present research results^{22, 19}.

Sprout length

Significant variations (p≤ 0.05) were found regarding sprout length as affected by various concentrations of growth regulators (Table-1). The treatments of IBA escalated with the maximum sprout length as compared to PHB and NAA registering the highest (107.20 cm) sprout length. Sprout length decreased at the highest IBA concentration of 5000 ppm with (102.53 cm).  PHB produced sprouts with lesser length than IBA and NAA which showed a decline at the higher concentrations beyond 750 ppm which showed the maximum (65.66 cm) sprout length. Augmentation of nutrient  availability might have  resulted in the plant metabolism as reported in ficus spp.²⁹ Increased linear growth of stem due to cell elongation by the action of auxins and maximum sprout length was recorded with the application of IBA and NAA as reported in scented geranium cuttings^30. Decline in the shoot length by PHB treatments has also been reported which are in sustenance with the present findings^{31, 32}.

Shoot weight (Fresh and dry)

Maximum (56.30 g) fresh weight was depicted by (IBA 4000 ppm).It was followed by PHB which also showed an increase up to 750 ppm with 47.00 g showing decline at higher concentrations but a great decline was noticed with PHB 1500 ppm accounting to 30.00 g of shoot fresh weight (Table-1). There was an increase in the shoot weight to a certain extent in NAA treated cuttings respectively. The untreated cuttings surpassed the PHB concentration of 1500 ppm weighing 35.63g as shoot fresh weight. The shoot dry weight followed the same trend with the maximum (12.19 g) observed in (2000 ppm IBA). The treatments of NAA was followed by IBA with the highest (9.18 g) at NAA 300 ppm. Among PHB treatments the shoot dry weight decreased below 750 ppm from 8.50 g to 4.71 g at 1500 ppm which was the least of all the treatments but the untreated cuttings was greater than this treatment .The increased weight with IBA can be attributed to the role of auxins in increasing the cell permeability for moisture and essential nutrients resulting in the cell division and its enlargement leading to the plant growth in terms of shoot number and their higher fresh shoot weight. Similar results are confirmed in fig³³,  dragon fruit cuttings ^{19, 22, 17.} The decline in the shoot weight at higher concentrations of growth regulators might be due to the toxicity of K+ ions³⁴. The low response of PHB than IBA can be attributed to its inhibition action as reported in peach³⁵. The superiority of IBA over NAA has also been reported in dragon fruit cuttings¹.

Survival percentage

According to the results incurred  72% of cuttings treated with the growth regulators survived (Table-1). Out of applied growth regulators the treatments which were treated with IBA prompted to the best in plant survival with maximum (90.7%) in the cuttings treated with IBA 4000ppm significantly. Minimum survival (32.70) was achieved in the cuttings treated with PHB 1500ppm. Distinct differences in survival rate of plants raised from cuttings with the application of growth regulators was noticed. PGRs such as IBA and NAA accelerates the rooting rate resulting in the increase of survivability of cuttings^{36, 37}. IBA proved to be the best auxin for general use due to its non-toxicity to plants than NAA³⁸. Its effective role in rooting and survival promotion has been advocated³⁹. The research findings in Kiwi fruit⁴⁰, pomegranate⁴¹ and  dragon fruit advocated the highest survival with IBA ^{13, 42, 17}.

Number of roots per cutting

Significantly (p≤ 0.05) increase in the root number was observed with (21.56) roots produced with the 4000 ppm IBA application. The treatments of NAA followed IBA with 18.54 roots being the maximum with NAA 300 ppm. PHB lagged behind in the root production than IBA and PHB with the root production range of 16.15 -12.16 being the highest with 750 ppm and the lowest at the highest concentration of 1500 ppm. Untreated cuttings performed better than PHB 1500 ppm with the root production of 12.99 roots per cutting respectively. More root production with IBA pertains to the promotion of cell division and their elongation with the auxins resulting in distinctness of cambium  formation into root initials due to the  availableness of  the energy reserves thus producing more roots⁴³. A correlation between primordial division in root initiation and endogenous or exogenous auxin results in the increase of roots ⁴⁴. The increase in the degree of root formation in cuttings with increasing IBA concentration has been reported in dragon fruit cuttings^{45, 1}. There was a reverse tendency in root production pertaining to  IBA dosage. These results are in conformity with in fig³⁰. Lesser root production with PHB than IBA has also been reported in pomegranate ³², fig ³¹,  peach ³⁵ and in olive⁴⁶.

Average length of root

Significantly (7.63cm) as maximum length of root was recorded in IBA 4000ppm. Out of NAA and PHB highest (6.10 cm) was recorded in NAA 300ppm (Table-2). Reduction in the root length at the highest concentration was reported in all the growth regulators tried. The research findings described that the increment in root length can be attributed to high C/N ratio, sucrose and reducing sugars produced as a result of enhanced carbohydrates disintegration depleting starch, their accumulation at the action site which in turn synthesise new proteins causing cell division and elongation^{17, 22}. These factors shows their correlation with enhanced rooting and its further growth in dragon fruit⁴. Decreased parameters at  the highest growth regulator dosage might have produced toxicity  in  the  cuttings³⁴.

Longest root length

According to the results depicted in (Table-2) significantly longest of all the roots (14.50 cm) length was recorded in (IBA 4000 ppm). On the other hand, out of NAA the longest root was of the length (10.10 cm) recorded in (NAA 500 ppm). PHB generated the longest root with length showing a great reduction at the highest concentration of 1500 ppm with only 1.00 cm which was lesser than the untreated cuttings with 3.36 cm respectively. The longest root production with IBA might be due to an increased metabolic activity of hormones in young tissues, availability of essential nutrients, balanced C:N ratio and better utilization of higher carbohydrates⁴⁷ . The present findings are akin to the results in dragon fruit^{17, 22}.

Root weight

The highest  (6.13 and 3.01g) fresh and dry weight was in (4000 ppm) IBA treated cuttings and the lowest in PHB-1500ppm (Table-2). The treatment of PHB on cuttings produced lower fresh weight of roots compared to IBA. The treatment of PHB 1500 ppm proved to be the poor in generating weight accounting to 0.50 g which was even lesser than the untreated cuttings with 0.70 g and 0.55 g of fresh  and dry weight respectively. The role of the auxins naturally occurring or exogenously applied might be the reason for root initiation and its invigoration.  Increased root dry weight with improved root traits with IBA due to the  higher dry matter accumulation has been ascribed in dragon fruit¹⁷. Heaviest roots with enough dry weight with IBA than PHB has also been reported in fig³¹. Decreased shoot weight has been attributable to the  toxic effect of K+ ions produced in the cuttings³⁴.

Table 1: Sprouting and vegetative growth  in dragon fruit cuttings as affected by plant growth regulators

Treatments	Number of Sprouts	Shoot emergence	Survival percentage	Length of shoot  (cm)	Fresh shoot weight(g)	Dry shoot weight(g)
T1 -NAA 100ppm	1.48±0.08j	53.31±2.06j	72.80±0.30i	56.07±1.36l	37.93±0.20l	6.71±0.15k
T2 -NAA 200ppm	1.72±0.05gh	57.96±4.12i	78.10±0.30f	71.60±2.43e	39.03±0.41k	7.15±0.20j
T3 – NAA 300ppm	1.98±0.03e	63.02±3.29g	82.00±0.30d	79.40±2.33d	45.50±0.20f	9.18±0.20e
T4 -NAA 400ppm	1.63±0.12hi	67.33±4.96e	83.50±0.30c	69.53±1.32f	41.63±0.41i	8.87±0.20f
T5 -NAA 500ppm	1.11±0.08l	65.15±4.23f	71.20±0.30j	62.60±3.75i	40.33±0.41j	8.19 ±0.20h
T6 -IBA 1000ppm	1.78±0.07fg	60.82±4.49h	74.40±0.30h	59.10±1.21jk	48.74±0.20d	8.43±0.20g
T7 -IBA 2000ppm	2.12±0.07d	70.44±3.63d	80.50±0.30e	67.66±2.88g	49.66±0.35d	9.96±0.20c
T8 -IBA 3000ppm	2.54±0.10c	74.54±5.09c	84.60±0.30c	93.26±1.85c	50.73±0.41c	9.40±0.20d
T9 -IBA 4000ppm	2.94±0.05a	79.27±4.80a	90.70±1.47a	107.20±1.44a	56.30±0.80a	12.19±0.20a
T10 -IBA 5000ppm	2.74±0.06b	77.02±4.46b	70.36±0.85k	102.53±2.40b	52.80±0.80b	10.53±0.20b
T11 -PHB 500ppm	1.53±0.05ij	39.78±3.56m	87.50±0.30b	57.93±0.95k	36.70±0.26m	6.20±0.20m
T12 -PHB 750ppm	1.86±0.11ef	47.79±4.96k	75.60±0.30g	65.66±0.76h	47.00±0.20e	8.20±0.20h
T13 -PHB 1000ppm	1.35±0.09k	51.41±2.91j	55.70±0.30l	60.60±0.72j	44.30±0.20g	7.59±0.20i
T14 -PHB 1250ppm	1.11±0.09lm	43.57±6.51l	45.60±0.30m	53.66±0.76m	43.10±0.20h	6.44±0.20l
T­15 -PHB 1500ppm	0.99±0.07m	42.09±5.52l	32.70±1.99n	40.33±0.57o	30.00±2.00o	4.71±0.48o
T16 -Control (Distilled water)	0.83±0.08n	32.90±4.98n	65.16±0.30k	49.33±0.76n	35.63±0.91n	5.70±0.20n
Mean ± S.D	1.73±0.61	57.90±14.34	71.90±15.31	68.53±18.35	43.71±6.89	8.09±1.90
CD (p≤ 0.05)	0.13	1.98	1.19	1.79	1.06	0.19

Numbers in rows and columns followed by the same letters show no difference.

Table 2: Rooting  in dragon fruit cuttings as affected by plant growth regulators

Treatments	Root number	Root length (cm)	Longest root (cm)	Fresh root weight (g)	Dry root weight (g)
T1 -NAA 100ppm	15.44±0.40h	3.70±0.20h	5.30±0.20j	1.20±0.20k	0.84±0.20k
T2 -NAA 200ppm	17.91±0.13e	4.03±0.20g	7.43±0.25h	2.00±0.20i	1.25±0.20i
T3 – NAA 300ppm	18.54±0.40d	6.10±0.20c	10.10±0.20d	3.50±0.20d	2.13±0.20d
T4 -NAA 400ppm	16.01±0.37g	5.30±0.20d	9.60±0.20e	2.90±0.20e	2.00±0.20e
T5 -NAA 500ppm	15.03±0.17i	4.50±0.20f	8.70±0.20f	2.30±0.20gh	1.60±0.20h
T6 -IBA 1000ppm	17.21±0.16f	3.10±0.20j	5.40±0.20j	0.80±0.20m	0.65±0.20lm
T7 -IBA 2000ppm	18.29±0.20d	3.40±0.20i	6.40±0.20i	1.00±0.20l	0.70±0.20l
T8 -IBA 3000ppm	19.39±0.15c	5.51±0.15d	10.63±0.42c	4.20±0.20c	2.48±0.20c
T9 -IBA 4000ppm	21.56±0.20a	7.63±0.25a	14.50±0.20a	6.13±0.11a	3.01±0.11a
T10 -IBA 5000ppm	20.46±0.12b	6.63±0.25b	13.20±0.20b	5.10±0.20b	2.70±0.20b
T11 -PHB 500ppm	14.91±0.13j	2.70±0.20k	4.20±0.20k	1.50±0.20j	0.97±0.20j
T12 -PHB 750ppm	16.15±0.15g	4.90±0.20e	8.96±0.11f	2.60±0.20f	1.86±0.15f
T13 -PHB 1000ppm	15.02±0.16i	4.23±0.25g	8.20±0.20g	2.40±0.20g	1.72±0.20g
T14 -PHB 1250ppm	14.10±0.10j	3.20±0.20ij	6.10±0.20i	2.20±0.20h	1.08±0.12j
T­15 -PHB 1500ppm	12.16±0.08l	0.59±0.45 m	1.00±0.50m	0.50±0.20n	0.25±0.13n
T16 -Control (Distilled water)	12.99±0.10k	1.55±0.47l	3.36±0.75l	0.70±0.20m	0.55±0.20m
Mean ± S.D	16.53±2.62	4.18±1.81	7.70 ±3.50	2.43±1.59	1.48±0.82
CD (p≤ 0.05)	0.36	0.25	0.437	0.17	0.12

Numbers in rows and columns followed by the same letters show no difference

Conclusion

The present study divulged the substantial potential of plant growth regulators leading to alterations in the shoot induction and root system with improved agronomic parameters. The hitherto conducted investigation revealed the effect of 4000 ppm IBA in dragon fruit cuttings for raising good quality plants. The conclusion of the research is in the context of the future application of stem cutting with IBA 4000 ppm as a means of the multiplication of the superior plants  on commercial basis.

Acknowledgment

The authors are thankful to the Department of Horticulture, Khalsa College, Amritsar, India for the help and support rendered.

Funding Sources

The authors received no financial support for this research.

Conflict of Interest

The authors declare no conflict of interest. 

Author^,s Contribution

Conceptualization of research work and Designing of work (AK) ; Execution of field/Lab experiments and execution of data (DS) ; Analysis of data and interpretation (AK and DS) ; Preparation of manuscript (AK); Both authors read and approved the final manuscript. 

Data Availability Statement

Not applicable 

Ethics Approval Statement

The study  did not involve  an experiment on humans and animals.

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