Cluster Bean (Cyamopsis tetragonoloba L.) Germination and Seedling Growth as Influenced by Seed Invigoration Techniques

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1 American-Eurasian J. Agric. & Environ. Sci., 15 (2): , 2015 ISSN IDOSI Publications, 2015 DOI: /idosi.aejaes Cluster Bean (Cyamopsis tetragonoloba L.) Germination and Seedling Growth as Influenced by Seed Invigoration Techniques Muhammad Aamir Iqbal Department of Agronomy, Faculty of Agriculture, University of Agriculture Faisalabad-38040, Pakistan Abstract: To investigate the comparative efficacy of different priming techniques and their effect on germination as well as seedling growth and development of cluster bean, a laboratory trial was conducted in the Allelopathy Laboratory of Department of Agronomy, University of Agriculture Faisalabad Pakistan, during The experimental design was complete randomized design (CRD) and was replicated four times. Eight seeds of cluster bean were placed in each petri dish. Cluster bean seeds were subjected to hydro priming, on-farm priming, halo priming with KNO 3 and priming with two concentrations (2% and 5%) of Moringa leaf extract as experimental treatments. A control treatment was kept for comparison. The results revealed that all priming techniques were effective in increasing the germination as well as growth and development of cluster bean. Priming with 5% Moringa leaf extract was instrumental in decreasing time to start germination (1.8 days) and time taken to 50% germination (3.1 days). It also increased final germination percentage (90%) along with better root and shoot length. Priming with 5% Moringa leaf extract also gave significantly higher root and shoot fresh weight (4.4 and 4.8 g respectively) and it was followed by 2% Moringa leaf extract. Hydro priming performed better than on-farm priming and halo priming with KNO 3 but was much less significant than either concentration of Moringa leaf extract. Key words: Allelopathy Halo priming Hydro priming Moringa leaf juice On-farm priming INTRODUCTION 1 cereal forages give less forage yield (50-60 t ha ) as compared to other forages such as berseem which is Pakistan continues to remain a country with an called as king of forages because of high forage yield as agriculture base as this sector contributes 21.4% share to well as quality. Cluster bean locally known as guar is an national gross domestic product (GDP) and more than annual, summer and drought tolerant leguminous crop 11% of this agricultural GDP is shared by livestock sector. with origin being the Indo-Pakistan subcontinent and best Pakistan is endowed with 37 million cattle, 32 million adapted to tropical and subtropical regions of the world. buffaloes, 28 million sheep and 68 million goat Pakistan ranks second among cluster bean growing populations with 51 million tons of milk production countries of the world [3]. It is grown as forage as well for annually [1, 2]. Livestock productivity and performance is extraction of guar gum (galactomannan) which finds its directly dependent upon the availability of forages utilization in food industry, a substitute of fat in human throughout the year. During spring season, mostly food, paper, pharmaceutical, petroleum and cosmetic sorghum and millets are grown as forage crops while in industry [4]. After gum extraction, the residue called autumn season; berseem fulfills the majority of forage guar meal is used a high protein concentrate for requirement in addition to forage oat. But the matter of animals [5]. The intercropping of cereal forages with concern is that cereal forages such as maize, sorghum, forage legumes such as cluster bean has been found millets and oat are poor in quality attributes particularly effective in increasing not only forage yield but also the protein contents as compared to other forages as berseem quality of mixed forage especially protein contents [6-15]. and alfalfa which result in suboptimal performance of Different researches have brought to light the fact that livestock, particularly the milch animals. Furthermore, forage legumes grown as an intercrop give less forage Corresponding Author: Muhammad Aamir Iqbal, Department of Agronomy, Faculty of Agriculture, University of Agriculture Faisalabad-38040, Pakistan. 197

2 yield as compared to their sole crop [16-18]. A variety of University of Agriculture Faisalabad Pakistan, during factors responsible for comparatively less forage yield of The experimental design was complete legumes in intercropping systems have been described randomized design (CRD) and was replicated four times. but poor germination and seedling establishment are Eight seeds of cluster bean (cv. BR-99) were placed in considered to be the major factors in reducing fresh each petri dish. Hydro priming was done by soaking forage yield [19, 20]. Different priming techniques have cluster bean seeds in distilled water for 12 hours and then been found to be effective in increasing crops germination shade dried before putting them into petri dishes and and seedling establishment in almost all field crops similarly on-farm priming was done in tap water for 12 [21-25]. Seed priming is pre-sowing strategy to affect hours. Halo-priming with 1% KNO 3 was done for 12 the pre-germination metabolic activities to enhance the hours. 2% and 5% solutions of Moringa leaf extract were germination [26-30]. Seed priming is done with different prepared and cluster bean seeds were soaked for 12 agents like tap water, distilled water, different salts and hours. Time to start germination (days), time taken to 50% other nutrients and vitamins. Seed priming improves germination (days), final germiation (%), root length (cm), germination by early DNA replication, increases RNA shoot length (cm), root fresh weight (g) and shoot fresh and protein synthesis, accelerates embryo growth, repairs weight (f) were recorded during the course of this the damaged and deteriorated seeds and causes a laboratory trial. The experiment was visited daily and the significant reduction in leakage of metabolites and finally time to start emegence, when first seed was germinated results in better germination, seedling establishment and was recorded. Time taken to 50% emergence was recorded better yield of field crops [31-38]. Hydro priming is simply by following the procedure suggested by Farooq et al. soaking of seeds in distilled water for duration (mostly 12- [50]. 24 hours) and then drying the seeds in shade before sowing. On-farm priming involves seeds dipping in tap E50 = t i+ (N/2 - n i) (t j- t i) (n j- n i) water for hours and then drying in shade before sowing [39]. Halo-priming is the technique of soaking Where N is the final number of emerged seeds, niand nj seeds in salt solutions containing different salts like KCl, are number of emerged seeds at times tiand tj. KNO 3, NaCl etc. [40-44]. Similarly, seed soaking in solutions containing different nutrient, plant growth Protocol for Moringa Leaf Extract (MLE) Preparation: regulators and promoters as well as vitamins, has also Moringa leaf extract (MLE) was prepared by collecting been found effective in increasing not only germination young and disease free leaves from Moringa tree. Theses and seedling stand but also significantly higher crops leaves were washed and then frozen for two days in yield were obtained. Similarly, Moringa (Moringa oleifera refrigerator at 4 C. Leaves were grinded in a manual juicer Lam.) tree has been recognized as a miracle tree as its leaf to extract the leaf juice. The juice was collected and extract contains sufficient quantities of macro and micro filtered by passing through a muslin cloth to remove all nutrients, particularly a substance called zeatin which has the green matter. After that the extract was stored at room the potential to increase crops yield when applied as a temperature. foliar spray. However Moringa leaf extract potential as a priming agent in increasing crops germination and Statistical Analysis: All the data collected were seedling establishment has not been explored [45-49]. subjected to ANOVA technique with the help of The present investigation was aimed to find out the MSTAT-C computer software program [51] and the least comparative efficacy and efficiency of different priming significant difference test at 5% probability level was agents and techniques in increasing the germination as employed to compare treatment means [52]. well as seedling growth and development of cluster bean. RESULTS AND DISCUSSION MATERIALS AND METHODS Time Taken to Start Germination (Days) and 50% To assess the comparative efficacy of different Germination (Days): Time taken to emergence depicts the priming techniques in increasing the germination and vigor of seeds as more vigorous seed emerge more rapidly seedling growth and development of cluster bean, a and take less time to start germination when soil and laboratory trial was designed and executed in the environmental conditions are conductive to germination. Allelopathy Laboratory of Department of Agronomy, The minimum time taken to start emergence (1.81 days) 198

3 Fig 1: Time taken to start germination (days) of cluster bean as influenced by different priming techniques. Fig 2: Time taken to 50% germination (days) of cluster bean as influenced by different priming techniques. was recorded by 5% Moringa leaf extract (MLE) treated better than control but these were much less seeds and it was followed by 2% Moringa leaf extract significant than priming with 5% Moringa leaf extract. (MLE) (1.96 days) (Fig. 1). The maximum time to start The significantly less time taken to 50% emergence was emergence (2.4 days) was taken by seeds that were not might be due to better metabolic activity within the seeds subjected to any priming technique (control). Even hydro that were treated Moringa leaf extract (MLE). These priming and halo priming performed better than control results are in line with Gniazdowska [53], Aregheore [54] but it was much less significant than priming with 5% and Khan et al. [55], who concluded that Moringa leaf Moringa leaf extract. This was might be due to growth extract has the potential to increase the rate germination promoters present in the Moringa leaf extract which might process by increasing the metabolic activity of seeds. had increased metabolic activity and ultimately germination started earlier than other treatments. The time Final Germination: Final germination shows the number taken to 50% emergence shows the time when half of of seeds that have been germinated out of total sown the seeds have been germinated. The minimum time seeds. Fig. 3 revealed that seed priming with 5% Moringa taken to 50% emergence (3.16 days) was recorded by 5% leaf extract (MLE) was instrumental in increasing the final Moringa leaf extract (MLE) treated seeds and it was emergence of soybean as it recorded the maximum final followed by seeds that were treated with 2% Moringa leaf germination (90%) and it was followed by 2% Moringa extract (MLE) (3.3 days) (Fig. 2). The maximum time to 50% leaf extract for 12 hours (84.5%). The minimum final emergence (3.8 days) was taken by seeds that were not germination was given by control treatment (66%). subjected to any priming technique (control) as even on- The significantly higher final germination given by 5% farm priming, hydro priming and halo priming performed Moringa leaf extract was might be due to zeatin which 199

4 Fig 3: Final germination (%) of cluster bean as influenced by different priming techniques. Fig 4: Root and shoot length (cm) of cluster bean as influenced by different priming techniques. is a natural plant growth regulator and other nutrients present in Moringa leaf extract. These results are in complete confirmation with Yasmeen et al. [56] and Foidle et al. [57], who observed more germination and seedling growth and development caused by zeatin. as well as macro and micro nutrients in Moringa leaf extract. These results are in line with Akinbode and Ikotun [58] and Phiri [59], who found Moringa leaf extract quite effective in increasing the seedling growth and development of many vegetable and agricultural crops. Root and Shoot Length (cm): Seedling growth and Root and Shoot Fresh Weight (g): Figure 5 reveals that development determines the final yield as good seedling the highest root weight (4.45 g) was recorded by 5% establishment constitutes the first and foremost step in Moringa leaf extract (MLE) priming and it was followed by achieving the full potential of crops. The maximum root 2% Moringa leaf extract (4.4 g). Similarly the highest shoot length (23.2 cm) was given by 5% Moringa leaf extract weight (4.8 g) was again observed in priming with 5% priming and it was followed by 2% Moringa leaf extract Moringa leaf extract and it was followed by 2% Moringa (22.6 cm) (Fig. 4). The minimum root length (17.34 cm) was leaf extract (4.7 g). The significantly higher root and shoot recorded by control treatment that was much less than all weight given by Moringa leaf extract was might be due other priming techniques. Similarly the highest shoot the action of zeatin which is a natural growth promoting length (24.5 cm) was given by seeds that were treated with substance belonging to the class of cytokinins. 5% Moringa leaf extract and it was followed by 2% These findings are in accordance with those reportd Moringa leaf extract (23.9 cm) (Fig. 4). The significantly by Phiri and Mbewe [60], Makkar and Becker [61] and higher root length recorded by Moringa leaf extract was Anjorin et al. [62], who found zeatin effective in might be due to the presence of various growth promoters increasing crops growth and yield. 200

5 Fig 5: Root and shoot fresh weight (g) of cluster bean as influenced by different priming techniques. CONCLUSION 5. Ayub, M., M. Khalid, M. Tariq, M.A. Nadeem and M. Naeem, Effect of different seeding Cluster been seed priming with 5% Moringa leaf densities and nitrogen levels on growth, forage extract has the potential to not only increase the yield and quality attributes of cluster bean germination but seedling growth and development (Cyamopsis tetragonoloba Tuab). J. Agric. Technol., of cluster bean is also improved and that too in a 7(5): cost-effective way as Moringa leaf extract is quite cheap 6. Addo, Q.A., A. Darkwa and G.K. Ocloo, and its preparation is also simple and easy for farmers. Growth analysis of component crops in a maize- It is also worth mentioning that Moringa leaf extract is soybean intercropping system as affected by time of environment friendly to its best and there is a need of planting and spatial arrangement. J. Agric. & Biol. more research on Moringa leaf extract regarding its Sci., 6(6): concentration as well as priming timings to develop it as 7. Ahmad, A.H., R. Ahmad, N. Ahmad and A. Tanveer, a future priming tool, particularly in the wake of rising Performance of forage sorghum intercropped input prices and skyrocketing population in millions that with forage legumes under different planting we will need to feed in times to come. patterns. Pak. J. Bot., 39: Ahmad, G., Z. Qureshi, D.S. Khan and A. Iqbal, REFERENCES Study on the intercropping of soybean with maize. Sarhad J. Agric., 17(2): Economic Survey of Pakistan, Govt. of 9. Ahmad, A.H., R. Ahmad and N. Mahmood, Pakistan, Ministry of Food, Agric. and livestock Div. Production potential and quality of mixed sorghum Economic Wing, Islamabad. forage under different intercropping systems and 2. Punjab development statistics, Govt. of Punjab, planting patterns. Pak. J. Agric. Sci., 44(2): Pakistan. Annual reports, pp: Arshad, M. and S.L. Ranamukhaarachchi, Ayub, M., M.A. Nadeem, M. Naeem, M. Tahir, Effects of legume type, planting pattern and time of M. Tariq and W. Ahmad, Effect of different establishment on growth and yield of sweet levels of P and K on growth, forage yield and quality sorghum-legume intercropping. Aus. J. Crop Sci., of cluster bean (Cyamopsis tetragonoloba L.). J. 6(8): Anim. Plant Sci., 22(2): Ayisi, K.K., M.S. Mpori and J. Vandenberg, Sortino, O. and F. Gresta, Growth and yield Grain yield response and chilopartillus infestation performance of five guar cultivars in a Mediterranean diverse sorghum crop intercrop management. South environment. Ital. J. Agron., 4: Agric. J. Plant Soil., 18:

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