Influence of different growing media and IBA concentration on rooting of cutting in mulberry (Morus alba L.)

Introduction

Over thousands of years, the domestication of Mulberry (Morus alba) has occurred, resulting in its adaptation to various tropical, subtropical and temperate zones across Asia, m Europe, North and South America and Africa (Ozgen et al., 2009) [20] . There are around 68 species of mulberry (Morus) worldwide, with Morus alba and Morus indica being the most prevalent types (Datta, 2000) [5] . The primary use of mulberry globally is for its leaves in the cultivation of silkworms, with China and India being the most significant countries in this regard. The Moraceae family encompasses Morus alba, and the majority of the native variants in India belong to the Morus indica genus. To reproduce fruit, it is recommended to utilize cultivated mulberry clones. Unlike tissue culture propagation and layering systems commonly used in plant production, grafting and cutting propagation techniques are typically employed (Guo et al., 2007) [7] . Among the various methods of vegetative multiplication, the use of hardwood cuttings is considered one of the simplest and most cost-effective. Recently, auxin treatments have been employed to stimulate the development of roots in cuttings (Fourrier, 1984) [6] . Several factors can influence the rooting potential of stem cuttings, including the specific requirements of the species and cultivar, the source and type of cutting, the presence of wounds or removal of leaves, etiolation of the stock plant, planting time and growing conditions such as the growth medium, misting, bottom heat, use of hormones, fertilizer and supplemental lighting (Hartmann et al., 2002) [11] . In comparison to studies on softwood cuttings, a considerable number of studies on mulberry have predominantly focused on hardwood cuttings and have been conducted on a large scale. In addition, the selection of a suitable rooting medium plays a crucial role in the growth and advancement of plants obtained from stem cuttings (Bakhsh et al., 2000; Yıldız and Koyuncu 2000; Koyuncu et al., 2004) [3,18] . The propagation process necessitates the inclusion of rooting media as a vital component. A recommended combination of peat, perlite, and coco peat in a ratio of 2:2:1 has been found to promote taller plant growth and enhance leaf production in citrus cuttings. Furthermore, successful results have been achieved by utilizing mixtures comprising vermiculite, peat, and perlite Albouyeh (2007) [1] . The auxin hormone, which is wellknown for its role in stimulating plant growth and the development of leaves, fruits, shoots, and lateral roots, is employed in stem cutting techniques. In a study conducted by Sokhuma et al., (2018) [28] , IBA and NAA, two specific types of auxin, were utilized as tools for stem cuttings. The researchers focused on the rooting and axillary branch outgrowth of mulberry stem cuttings. Interestingly, they discovered that the application of 1,000 mg of IBA and NAA did not have any noticeable impact on the percentage of stem cuttings that successfully rooted (Sokhuma et al., 2018) [28] . Mulberry is commonly cultivated through the use of hard wood cuttings due to its various advantages, such as the rapid multiplication of parent material and the ability to maintain desirable plant properties. The timing of cutting preparation plays a crucial role in determining the quantity and success of root development in mulberry. Factors such as the physiological state of the plant and its surrounding environment influence the optimal time for cutting preparation and planting. Additionally, the current weather conditions and outside temperature impact the collection of cuttings and their subsequent rooting success. Other factors that can affect the likelihood of stem cuttings rooting include the plant's juvenility and overall condition, as well as the source, location, and type of cutting taken. Furthermore, any injury or leaf removal can also influence the rooting process. It is important to consider these factors in order to maximize the success of mulberry propagation through stem cuttings (Hartmann et al., 2002) [1] . The rooting ability of cuttings is influenced by various internal and external factors, including the type and concentration of hormones present in the cuttings, as well as the variety and planting season. In the context of large-scale mulberry plant reproduction, the application of growth regulators to cuttings has proven to be more advantageous and cost-effective compared to the grafting method (Rao and Khan, 1963) [24] . So considering all these, this study aims to evaluated the influence of different growing media and different concentration growth regulator on cutting of mulberry.

Materials and Methods

The study "Influence of different growing media and IBA on rooting of cutting in mulberry (Morus alba L)" was conducted in the experiment farm of school of agriculture, department of Horticulture, Lovely professional University under shade net house during the academic year (2023-2024). The variety used for this trial was collected from the field of agriculture farm lovely professional university, Punjab. The main objectives of study the effect of growth regulator on rooting of cutting and influence of different growing media on root and shoot growth of mulberry. The study was conducted using a completely randomized design, consisting of three replications and seven treatments. Each treatment included ten cuttings. One-year-old cuttings of the fruitless cultivar of white mulberry (Morus alba L) were acquired from mature and healthy trees in the gardens of the main campus of the University, using a sterilized wood cutter. Cuttings were made using partially mature branches with a thickness ranging from 0.75 to 1.00 cm. These cuttings, measuring 15-20 cm in length and containing 5-6 buds, were prepared for planting by completely removing the leaves. Various growth substrates such as soil, farmyard manure (FYM), cocopeat, vermiculite and perlite were prepared. Subsequently, different concentrations of IBA solution (1000 ppm, 2000 ppm, 3000 ppm) were prepared from the stock solution in the laboratory. The prepared substrates are then placed into polybags and transferred to the shade net house. The prepared cuttings were immersed in IBA solutions of varying concentrations before being placed into the polybags. The study focused on evaluating the effect of different growing media and IBA concentration on mulberry hardwood cuttings. A total of seven treatment combinations were prepared viz., T 1 (control), T 2 (Soil + FYM + IBA@1000ppm), T 3 (Soil + FYM + IBA@2000ppm), T 4 (Soil + FYM + IBA@3000 ppm), T 5 (Cocopeat + Vermiculite + Perlite + IBA@1000 ppm), T 6 (Cocopeat + Vermiculite + Perlite + IBA@2000 ppm), T 7 (Cocopeat + Vermiculite + Perlite + IBA@3000ppm). Various vegetative and root growth parameters were evaluated. After mulberry cuttings, were subjected in poly house, they were investigated in the way maximum number of days required for sprouting per cutting, the number of sprouts per cutting, the percentage of sprouted cuttings, the survival percentage of sprouted cuttings, the number of leaves, the length and width of the leaves in centimeters, the fresh weight and dry weight of the shoots in grams, the number of roots generated per cutting, the number of primary roots, the number of axillary roots, the length of the longest root in centimeters, as well as the fresh and dry weight of the roots in grams.

Results and Discussion

Goal of the experiment was to determine the effect of growth regulator on rooting of cutting and influence of different growing media on root and shoot growth of mulberry (Morus alba L). In addition to organic amendments like soil, vermiculite, perlite and cocopeat, the treatments included changes in the concentration of indole-3-butyric acid (IBA). As per table 1 treatment T 4 (Soil + FYM + IBA 3000 ppm) showed the shortest time, with cuttings sprouting (6.33) as compare to the other treatments followed by T 3 , T 2 , and T 7 . T 1 showed a more days-to-sprouting with a value of (15.33). After conducting a thorough analysis of various parameters, it was determined that the application of IBA at a concentration of 100 ppm is the most effective plant growth regulator for the vegetative propagation of Morus alba under the current environmental conditions (Rafeeq et al., 2020). The similar findings have also been reported by Khan et al., (2007) investigated the best Sprouting percentage and number of leaves was found maximum in T 4 . The data pertaining number of sprouts per cutting in T 4 having the maximum number of sprouts (30.33) as per table 1, followed by T 3 and T 2 . The minimum number of sprouts shown (14.33) in T 1 . Among all the treatments, numbers of sprouted cuttings, length of the roots per cutting, percentage of rooted cutting, lengths of longest root were higher in IBA 2000 mg /L investigated by (Singh et al., 2014) [26] . The similar findings have also been reported by Pallavi et al., (2018) [22] studied the effect of IBA and NAA on rooting and growth of mulberry cuttings. They found that maximum number of shoots (2.67) and number of leaves (7.31) was observed when treated with 2000 ppm IBA. The minimum value was observed when treated with control. The variation in dose response to the number of roots might be attributed to the varietal and climatic differences in the location. The sprouting percentage varied significantly between treatments, showing the varying impacts of soil amendments and hormone combinations on the rooting performance of mulberry cuttings. As per table 1 T 4 exhibited the highest sprouting rate (91.11%) than other treatments and T 1 showed the minimum sprouting percentage (35.55%). In case of rooting media, the maximum number of sprouted cutting, diameter of thickest sprout, number of leaves on new shoots, survival percentage, rooting percentage, number of primary root, length of longest root was observed under vermicompost rooting media by (Singh et al., 2018) [27] . The similar findings reported by Kalyoncu et al., (2009) [15] . Among the treatments, T 4 exhibited the highest survival percentage of sprouted cuttings, with a rate of (95.55%). In T 3 and T 2 survival percentages of were (77.78%) and (66.67%) respectively. On the other hand, the treatment T 1 exhibited lower survival percentage of (40.00%) compared to the other treatments. In terms of the survival percentage of sprouted cuttings in T 7 , T 6 , and T 5 showed moderate rates with IBA concentrations of 1000, 2000, and 3000 ppm. The role of indole-3-butyric acid (IBA) for rapid propagation of mulberry for higher biomass and large-scale production; and examines the associated biochemical changes during rooting. The non-treated (control) and treated (1000, 2000 and 3000 IBA mg L -1 ) soft stem cuttings were cultured in mist chamber; IBA increased rooting phenomenon and activated carbohydrate metabolism (Husen et al., 2015) [12] . similar finding also have been reported by Hawramee et al., (2019) [9] cuttings treated with different concentrations of IBA significantly increased success percentage. the highest values of success rate of rooting 1 st and immersed in 1500ppm of IBA solution (T 7 ), while success rate of rooting (58.33%) and all other qualitative parameters of shoots height were obtained from cuttings taken on the same time but treated with 2000 ppm. The number of leaves per cutting shown highest in T 4 with the value (25.33) consistently produced competitive or higher leaf counts (25.33) at 90 days after planting, indicating that enhance leaf growth as shown in table 2. On the other hand, T 1 showed less number of leaves (11.33) under controlled conditions (Table 2). The study shows that the effect of IBA concentrations on rooting of mulberry cuttings with varying number of buds, the data of two budded cuttings treated with 3000 ppm of IBA showed at par results with three budded cuttings treated with 3000 ppm of IBA on rooting parameters and also proved that IBA play a significant role in rooting of mulberry saplings (Pooja et al., 2022) [8] . The similar findings have also been reported by Karabulut and Saracoglu (2022) [16] , investigated the impact of IBA and cinnamic acid treatments on the rooting of black mulberry cuttings. The results revealed that among the different treatments and periods, the combination of 6000 ppm IBA + 100 ppm CA applied during July and January exhibited superior performance compared to the others. 134 Treatment combination: T1 (control), T2 (Soil + FYM + IBA@1000), T3 (Soil + FYM + IBA@2000), T4 (Soil + FYM + IBA@3000), T5 (Cocopeat + Vermiculite + Perlite + IBA@1000), T6 (Cocopeat + Vermiculite + Perlite + IBA@2000), T7(Cocopeat + Vermiculite + Perlite + IBA@3000) 122 Treatment combination: T1 (control), T2 (Soil + FYM + IBA@1000), T3 (Soil + FYM + IBA@2000), T4 (Soil + FYM + IBA@3000), T5 (Cocopeat + Vermiculite + Perlite + IBA@1000), T6 (Cocopeat + Vermiculite + Perlite + IBA@2000), T7(Cocopeat + Vermiculite + Perlite + IBA@3000)

The length of the leaves per cutting was significantly affected by the concentrations of IBA and the various soilless media used. The longest leaf length of (24.00 cm) was observed in T 4 as shown in table 2. It was statistically at par with treatments T 3 and T 2 (Soil +FYM) with different concentration of IBA (1000, 2000ppm) having value (21.33 cm), (18.00 cm). On the other hand, the smallest leaf length of (8.66 cm) was observed in T 1 . The study's findings demonstrate that the amount length of leaves was significantly increased by applying 3000 ppm IBA. A similar result was found by Polat, (2008) [21] the effect of Indole butyric acid on rooting of mulberry Cuttings four local cultivars of mulberry were assessed for IBA rooting. One-yearold shoots were cut into 20-25 cm lengths throughout the dormant stage. Before planting, basal sections (1-2 cm) of the cutting were treated with IBA solution at different doses for 10 seconds. After 93-118 days of cutting, rooting proportion, root number per cutting, and average root length (cm) were measured. Cuttings treated with 5000 ppm IBA provided the best rooting performance. The data clearly indicates that both the IBA concentrations and various soilless media have a significant impact on the width of the leaves per cutting T 4 having the maximum width of the leaf, measuring (7.00cm) as shown in table 2. Conversely, the T 3 and T 2 treatments had narrower leaf widths compared to T 4 , with T 3 measuring (6.00cm) and T 2 measuring (5.22 cm). The control T 1 , displayed the minimum leaf width of (3.07cm) than all other treatments. Among the treatments, treatment T 4 exhibited the highest fresh shoot weight, measuring (37.03 g) T 4 treatment having the width of the leaf, measuring (7.00 cm) as shown in table 2. The control group (T 1 ) showed lower shoot weights compared to the all treatments. The treatment with the highest dry weight of shoot was observed in T 4 (9.44 g) followed by T 3 a dry shoot weight of (7.26g), T 2 produced a dry shoot weight of (6.25 g). Control (T 1 ) showed the lowest dry weight of shoot, measuring (2.54 g) then all other treatments as shown in table 2.: T1 (control), T2 (Soil + FYM + IBA@1000), T3 (Soil + FYM + IBA@2000), T4 (Soil + FYM + IBA@3000), T5 (Cocopeat + Vermiculite + Perlite + IBA@1000), T6 (Cocopeat + Vermiculite + Perlite + IBA@2000), T7(Cocopeat + Vermiculite + Perlite + IBA@3000)

As per table 3, T 4 exhibited the number of roots generated was the highest, with a value of (9.00) then all other treatments. T 1 had the lowest value (4) among all treatments. It is evident that the primary roots were most abundant in T 4 with value (9.60). Following closely behind were T 3 and T 2 respectively, with values of (7.83) and (6.19). On the other hand, the control group (T 1 ) had the lowest number of primary roots, measuring at (4.06). The number of roots generated was the highest in T 4 , with a value of (9.40) then all the other treatment. Whereas, T 5 with a concentration of IBA (1000 ppm) showed a value of (3.97), T 6 and T 7 values of (4.93) and (5.40) respectively. T 1 had the lowest value among all treatments (4.13). The data conferred table 3 revealed that the length of longest root was significantly influenced by the use of IBA concentrations and different soilless media. Maximum length of longest root (25.07 cm) was found in treatment T 4 . Treatment T 1 (control) recorded minimum average length of longest root (11.00 cm). T 3 and T 2 having the value (19.50 cm) and (18.00 cm). As per table 3, T 4 exhibited a significant rise in fresh root weight (7.42). Similarly, Treatment T 3 showed an elevated fresh root weight, reaching (6.85 g). This highlights the positive impact of using FYM organic amendments on enhancing root biomass accumulation, thereby promoting the robust growth of mulberry cuttings. On the other hand, Treatments T 5 , T 6 , and T 7 with varying IBA concentrations resulted in fresh root weights of (2.27g), (2.90 g), and (5.17g) respectively. In contrast, Treatment T 1 (control) exhibited the lowest fresh weight (2.70 g) compared to all other treatments. Treatment T 4 , resulted in a substantial enhancement in dry root weight, measuring (2.467g). Table 3 highlight that T 3 , which involved soil, FYM, and IBA at 3000 ppm, also contributed to an increase in dry root weight. T 5 , root weight (1.27 g), T 6 , and T 7 , resulted in fresh root weight (1.40 g), and (1.60 g), respectively. In contrast, T 1 (control) exhibited the lowest dry weight of (1.13 g) compared to all the other treatments. The effects of (IBA) dosages and relative humidity on softwood top cuttings of two varieties of black mulberry were examined. Cuttings were applied to the various IBA doses. The maximum root branching value was discovered from Type 3 in 3000 ppm IBA dose application (16.20 number per cutting), and the longest root was determined from 3000 ppm IBA dose of Type 1 (11.23 cm). In a study investigation of IBA at different dose on mulberry cutting (Morus alba) was conducted. Which showed that stem cuttings of the plant were subjected to different treatments; IBA concentration, IBA was found to be most effective at 4000 ppm for best rooting and good root growth, and at 2000 ppm for healthy leaf growth and to enhance the number of primary and secondary roots in Mulberry (Regmi et al., 2022) [25] . The highest root numbers were obtained when 6000 ppm IBA and 2000 ppm Pacroburazol applications. Cekic et al., (2013) [4] . Similar studies also found by Singh et al., (2018) [27] , the effect of auxins and rooting media on rooting in stem cutting of mulberry (Morus nigra L.). They used the different treatments IBA concentration and rooting media (Sand+ FYM, Vermicompost and Coco peat). In case of IBA concentration, the maximum number of sprouted cutting, average length of longest sprout, length of longest root was recorded under 2000 ppm concentration of IBA. In case of rooting media, the maximum number of sprouted cutting, diameter of thickest sprout, number of leaves on new shoots, survival percentage, rooting percentage, number of primary root, length of longest root was observed under vermicompost rooting media. The basal portions of mulberry hardwood stem cuttings were subjected to different concentrations of IBA using the slow dip. The similar findings have also been reported by Aziz et al., (2020) [2] the effects of different concentrations of IBA and soaking in water for 24 hours on propagation black mulberry (Morus nigra L.) by hardwood cuttings. Effect of individual factors showed that the highest rooting percentage (15%) was achieved in cuttings soaked in water for 24 hours, as well as improving other traits. Also, the best (23.33%) rooting was found in cuttings dipped in 4000 ppm IBA. The findings revealed that the concentration of 3000 ppm of IBA yielded the most favorable results (Sokhuma et al., 2018) [28] .

Conclusion

Based on the findings of the current study, it can be concluded that the utilization of various treatments resulted in significant variations in several parameters. These parameters include the maximum number of days required for sprouting per cutting, the number of sprouts per cutting, the percentage of sprouted cuttings, the survival percentage of sprouted cuttings, the number of leaves, the length and width of the leaves in centimeters, the fresh weight and dry weight of the shoots in grams, the number of roots generated per cutting, the number of primary roots, the number of axillary roots, the length of the longest root in centimeters, as well as the fresh and dry weight of the roots in grams. Notably, treatment T 4 (Soil + FYM + IBA 3000 ppm) exhibited the highest values for all these parameters.

References

1. Aziz RR, Mohammed AA, Ahmad FK, Ali AJ, et al. Effect of IBA concentration and water soaking on rooting hardwood cuttings of black mulberry (Morus nigra L.). Journal of Zankoy Sulaimani -Part A (JZS-A). 2020;22(1):153-158.
2. Baksh S, Mir MR, Darzi GM, Khan MA, et al. Performance of hardwood stem cuttings of mulberry genotypes under temperate climatic conditions of Kashmir. Indian Journal of Sericulture. 2000;39(1):30-32.
3. Cekic C, Erdem SO, Aydemir M, et al. The effects of pacrobutrazol and IBA treatments on the rooting of woodcuttings of black mulberry (Morus nigra L.) and red mulberry (Morus rubra L.). TABAD, Tarım Bilimleri Araștırma Dergisi. 2013;6(1):174-177.
4. Datta RK. Mulberry cultivation and utilization in India. FAO Animal Production and Health Paper. 2002;147(1):4562.
5. Fourrier B. Hardwood cutting propagation at McKay nursery. In: Proceedings of the International Plant Propagators Society. 1984;34(1):540-543.
6. Wel-Guo Z, Tingting C, Yonghua Y, Yi-Le P. A preliminary analysis of asexual genetic variability in mulberry as revealed by ISSR markers. International Journal of Agriculture and Biology (Pakistan). 2007;9(6).
7. Pooja HS, Sadatulla F. Effect of IBA concentrations on rooting of mulberry cuttings with varying number of buds. The Pharma Innovation Journal. 2022;11(12):1912-1914.
8. Hawramee OA, Aziz RR, Hassan DA. Propagation of white mulberry (Morus alba L.) fruitless cultivar using different cutting times and IBA. In: IOP Conference Series: Earth and Environmental Science. 2019;388(1):012-069.
9. Husen A, Iqbal M, Siddiqui SN, Sohrab SS, Masresha G. Effect of indole-3-butyric acid on clonal propagation of mulberry (Morus alba L.) stem cuttings: rooting and associated biochemical changes. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences. 2017;87:161-166.
10. Kako SM. The effect of auxin IBA and kinetin in budding success percentage of mulberry (Morus sp.). International Journal of Pure and Applied Sciences and Technology. 2012;13(1):50.
11. Kalyoncu H, Ozer E. Gilaburu'nun (Viburnum opulus L.), green side köklendirilmesi steels and to obtain seedlings. In: Proceedings of the Second National Nursery Symposium. 2000;25-29.
12. Kalyoncu IH, Ersoy N, Yılmaz M, Aydın M. Effects of humidity level and IBA dose application on the softwood top cuttings of white mulberry (Morus alba L.) and black mulberry (Morus nigra L.) types. African Journal of Biotechnology. 2009;8(16).
13. Karabulut NS, Saraçoğlu O. The effects of cinnamic acid and IBA treatments on the rooting of wood cuttings of black mulberry (Morus nigra L.). Turkish Journal of Food and Agriculture Sciences. 2022;4(1):1-5.
14. Khan GS, Ahmad I, Nawaz MF. Sprouting percentage and growth behavior of Morus alba as affected by size of cuttings and polythene tunnel. Pakistan Journal of Agricultural Sciences. 2007;44(2):321-328.
15. Koyuncu F, Vural E, Çelik M. Kara dut (Morus nigra L.) çeliklerinin köklendirilmesi üzerine araştırmalar. In: Proceedings of the National Kiwi and Berry Fruits Symposium. 2004;424-427.
16. Hawramee OA, Aziz RR, Hassan DA. Propagation of white mulberry (Morus alba L.) fruitless cultivar using different cutting times and IBA. In: IOP Conference Series: Earth and Environmental Science. 2019;388(1):012-069.
17. Ozgen M, Serçe S, Kaya C. Phytochemical and antioxidant properties of anthocyanin-rich Morus nigra and Morus rubra fruits. Scientia Horticulturae. 2009;119(3):275-279.
18. Polat AA. Effect of indolebutyric acid on rooting of mulberry cuttings. In: XXVII International Horticultural Congress-IHC International Symposium on Endogenous and Exogenous Plant Bioregulators. 2006;774:351-354.
19. Pallavi D, Sharma GL, Naik EK. Effect of IBA and NAA on rooting and growth of mulberry cuttings. International Journal of Current Microbiology and Applied Sciences. 2018;7(11):305-308.
20. Rafeeq J, Mughal AH, Zaffar SN, Dutt V, Ahmad K, Raja T. Effect of IBA on rooting and growth of Morus alba shoot cuttings under temperate conditions of Kashmir. International Journal of Chemical Studies. 2020;8(4):38003802.
21. Rao LS, Khan AA. Vegetative propagation of Japanese mulberry varieties by use of growth regulators. Indian Journal of Sericulture. 1963;1(3):723.
22. Regmi S, Puri C, Shah K. IBA impact at different dose on mulberry cutting (Morus alba). Research in Agriculture Livestock and Fisheries. 2022;9(3):253-257.
23. Singh KK, Choudhary T, Kumar A. Effect of various concentrations of IBA and NAA on the rooting of stem cuttings of mulberry (Morus alba L.) under mist house condition in Garhwal hill region. Indian Journal of Hill Farming. 2014;27(1):74-77.
24. Singh KK. Effect of auxins and rooting media on rooting in stem cutting of mulberry (Morus nigra L.). The Pharma Innovation Journal. 2018;7(11):12-15.
25. Sokhuma P, Intorrathed S, Phonpakdee R. Effect of IBA and NAA on rooting and axillary shoot outgrowth of Himalayan mulberry stem cutting. International Journal of Agricultural Technology. 2018;14(7):1939-1948.
26. Yildiz K. A study on propagation of mulberry (Morus nigra L.) by hardwood cuttings. Derim (Turkey). 2000;17(3).
27. Zenginbal H, Demir T. Effects of some rhizobacteria and indole-3-butyric acid on rooting of black and white mulberry hardwood cuttings. The Journal of Animal & Plant Sciences. 2018;28(5):1426-1431.