Study of the Anatomical Growth Parameters of lentil Plant to Pre-soaked Seed treatment with Water Extract of Wolffia arrhiza

doi:10.21203/rs.3.rs-4337063/v1

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Study of the Anatomical Growth Parameters of lentil Plant to Pre-soaked Seed treatment with Water Extract of Wolffia arrhiza

https://doi.org/10.21203/rs.3.rs-4337063/v1

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Wolffia arrhiza is known as noxious weed and rated as one of the ten weeds growing country in South – East Asia, Commonly infesting natural impounded water in India. Studies have been made for their utilization in multiple ways in developing countries. Present study based on 24 hours pre-soaking seed treatment with control, 1,2 and 5% concentration of water extract of W. arrhiza for anatomical growth of stem of lentil plants variety DPL-15.

The observation of stem show that the anatomical growth parameters like average diameter of stem, average thickness of stele, number of vascular bundles per microscopic field, average diameter of Xylem vessels, average diameter of Phloem is stimulated in seed treated with water extract for 24 hours. 5% level of water extract of W. arrhiza was found best for diameter of stem, number of vascular bundles per-microscopic field, and average thickness of stele. However for average diameter of Xylem vessels and average diameter of Phloem of lentil stem was observed better at 2% level of water extract of W. arrhiza. Results are statistically analyzed at 5% error probability.

Wolffia arrhiza

Anatomical growth

thickness of stele

Hormones

The dynamic features of aquatic habitats in South-East Asia in general and India in particular have supported luxuriant growth of aquatic weeds. Studies have been made for their utilization in multiple ways in developing countries. Aquatic weeds are known to cause problems of varied nature like covering of impounded water, hindrance to fisheries, choking of flowing waters, suppressing growth of cultivated plants, making water imputable, increasing loss of water, creating disease problems, causing impediment to navigation and hindrance to aquatic sports. Besides, such problems they also supplement as sources of fodder, green manure and fish food. In view of the significance of noxious aquatic vegetation, a general consciousness among scientists working in different regions of South-East Asia was evolved to pinpoint problems associated with vast community of aquatic plants and explore possibilities of their utilization for benefit of mankind.

Literature excels in excellent experimental work of eminent scientists dealing with aquatic plants; still there are wide gaps in our knowledge to fully exploit W. arrhiza attributes likely to offer solution on an array of botanical quizzes. Evolutionary voyage of angiosperm vessel structure-function and its significance for early angiosperm success reported by Field and Wilson (2012).This interesting perspective has mated only a causal arbitrary and partial treatment during the works of various investigators in the past. Nothing is known towards application of W. arrhiza extracts with special reference to growth, development and yield of Lentil plants, and how its metabolism and morph-anatomical attributes could be altered for better adaptability of the crop to multiply growth and better quality yield. Hillman (1961) rightly pointed out that “while adequate coverage by all work with a particular group of plants must perforce touch most fields of botanical research, it is impossible to consider each of the problems in its general context, to do so would be to write an Encyclopedia.Costanza et al., (2014) pointed changes in the global value of ecosystem services.sssDuckweeds in landscapes of Kanpur, India and mapping wetlands inter-alias significance suggested (Tiwari, 2000, 2002). Effect of water deficit and micro nutrients foliar application on the productivity of wheat plant studied (Kassab et al., 2004). Holtta et al., (2013) reported scaling of Xylem and Phloem transport capacity and resource usage with tree size. Correlative studies on growth and metabolism of duckweed's reported by (Pandey, 1979). This syn-optically background presented here sufficiently pinpoints existing state of knowledge and suggests areas of research on W. arrhiza forming theme of present investigation. Present investigation deals with utilization of W. arrhiza plants with special reference to anatomical growth parameters of stem of lentil plants.

Studies on utilization of W. arrhiza in agriculture were made with special reference to their use for obtaining extracts containing growth substances. Lentil varieties D.P.L-15 were selected to study the effect. Genetically tested seeds were obtained from Indian Institute of Pulse Research, Kanpur. Seeds of approximately same size and weight were selected for experiment.

Wolffia arrhiza was chosen as experimental material because of its quantitative abundance and growth in the district. Experimental material grown in stock cultures as described earlier was used for preparation of W. arrhiza extracts.

The extractions of W. arrhiza as growth-promoting substances were suspended in water. Five milliliter of W. arrhiza by volume was taken and ground in a clean porcelain mortar with water. In case of water extract sufficient water was added to make it 100 ml to have a five percent extract. 1,2, and 5 percent extracts were made by further dilutions with distilled water. Fifty seeds were soaked in sterilized petridishes in different concentrations (1, 2 and 5 percent) of water extracts of W. arrhiza and distilled water (control) for 24 hours.

The effect of treatments was studied under field earthen pots conditions in the garden beds laid for specific purposes with dimensions of 8 feet in breadth and 10 feet in length. Each bed was sown with 3 rows containing 9 seeds spaced 25 cm in rows 60 cm apart. Thus, total number of plants grown in each bed was 27 out of which 25 were selected for observations. Two beds of each treatment and normal untreated control were laid to raise 50 replicates. Seeds of lentil variety D.P.L-15 were soaked 24 hrs in various concentrations of extracts. Necessary plant protection measures were adopted to keep crop free of weeds, insect, pest and diseases.

Material of stem was cut carefully from control and water extracts in variety D.P.L.-15 pre-soaking seed treatment for 24 hours. Applied treated and control plants were grown in garden beds. Stem pieces of 2 cm were being collected from 5 cm below the top of the plant. Such materials of each treatment were preserved in formalin aceto – alcohol containing a mixture of 70 percent ethyl alcohol 90 ml., glacial acetic acid 5 ml. and formalin 5 ml. The material was then dehydrated and was micro-tome using senior rotary micro-tome model MT 1090 A. Slides of materials prepared were stained in safranin and fast green following Johansen’s technique (1940). Observation on diameter of stem, thickness of stele, number of vascular bundles, diameter of Xylem vessels and Phloem in stem sections in each treatment were measured with Ocular and Stage micrometer.

I. Diameter of stem (µ) – Increase in water extract of W. arrhiza increases the diameter of stem of lentil up to 5% level of water extract.

As compare to control, all the levels of water extracts significantly increases the diameter of stem of lentil.

Increase in diameter of lentil was found significantly higher at 1% over control, 2 over 1, and 5 over 2% level of water extract. Maximum value for diameter of stem of lentil was observed at 5% water extract level of W. arrhiza.

II. Average thickness of stele (µ) – Increase in water extract level of W. arrhiza increases average thickness of stele in lentil up to 5% level of water extract.

As compare to control, all the levels of water extract increases average thickness of stele, significantly higher at 1% over control, 2 over 1 and 5 over 2% level of water extract. Maximum value of average thickness of stele in lentil was observed at 5% water extract level of W. arrhiza.

III. Number of Vascular bundles per-microscopic field – Increase in no. of Vascular bundles of lentil increases with the increase in water extract level of W. arrhiza up to 5% level as compare to control, all the levels of water extract increases no. of Vascular bundles was found equal at 1, 2 and 5% of water extracts. Maximum no. of Vascular bundles of lentil was observed at 1% water extract level of W. arrhiza.

IV. Average diameter of Xylem vessels (µ) – Increase in water extract of W. arrhiza increases average diameter of Xylem vessels of lentil stem up to 2% level, further in increase in water extract level decreases the average diameter or Xylem vessels.

As compare to control 2% level of water extract significantly increases the average diameter of Xylem vessels. The other level faces to reach significant differences. Increase in average diameter of Xylem vessels was found to be significantly higher 2% over control, however 1% over control and 5% over 2% face to show significant differences. Maximum average diameter of Xylem vessels of lentil of stem was observed at 2% level of water extract of W. arrhiza.

V. Average diameter of Phloem (µ) – Increase in average diameter of phloem increases with the increasing water extract level of W. arrhiza up to 2% level; further increase in water extract decreases the average diameter of Phloem. As compare to the control 1 and 5% level of water extract showed insignificant increase, however 2% water extract level showed significant increase in average diameter of Phloem. Maximum average diameter of Phloem in lentil was observed at 2% water extract level of W. arrhiza.

Based on information scattered in the literature (Hillman, 1961) and preliminary observations made (Shukla, Pandey and Shukla, 1973) perimeters of study set up as described earlier for present investigation, bore fruits and revealed interesting results. They provided a new dimension of importance to lemonades. Holtta et al., (2009) suggested linking Phloem function to structure: analysis with a coupled Xylem-Phloem transport model. Vessel diameter- stem diameter scaling across woody angiosperms and the ecological causes of Xylem diameter variation reported by Olson and Rosell (2013). Barrera et al.,(2019) reported functional adaptation of Begonia oxysperma and Begonia romosii Merr. (Begoniaceae) revealed through morphs-anatomical analyses. Petit and Crivellaro (2014) studied comparative axial widening of Phloem and Xylem conduits in small woody plants. Security of wetlands and its agricultural and socioeconomic significance reported by Tiwari et al., (2009). Nutritive value of Indian foods studied by (Gopalan et al., 2004). Studies on growth, periodicity, succession, moisture, content and biomass of duckweed's in Kanpur suggested by (Shukla and Pandey, 1979). Utilization of W. arrhiza extracts for improvement of dry-weight and number of nodules in lentil crop and hydrophytes growth promoting substances and its consequence on NPK contents in lentil plant leaves reported by (Tiwari, 2017, 2021). Schulz (1998) studied Phloem structure related to function. Present investigation has brought to knowledge facts of both academic and applied significance. The utility of duckweed's in obtaining extracts to be employed in agriculture has further multiplied their importance. A correlative discussion of observations made during present investigation and facts recorded elsewhere in the literature would provide a conceptual synthesis of subject matter. On the basis of increase of diameter of stem, thickness of stele, number of vascular bundles 5% water extract of W. arrhiza was found significant result. However for average diameter of Xylem vessels and average diameter of Phloem of lentil stem was observed better result at 2% water extract of W.arrhiza. Seed treatment of W. arrhiza is responsible for inhances average diameter of stem, average thickness of stele, number of vascular bundles per microscopic field, average diameter of xylem vessels, average diameter of phloem may be due to presence of some growth promoting substances along with minerals present in extract.

The result is in support of earlier findings of some other water extract on other crops. Such as moringa leaf extract improves growth, biochemical attributes, and productivity of late - sown quiona (Rashid et al. 2021). Abdalla (2013) also reported similar results that foliar application of moringa leaf extract improve the plant height, fresh and dry weights, photosynthetic rates, the amount of each of chlorophyll a and b, stomatal conductance, total proteins, ascorbic acid and plant growth hormones like auxin, gibberillins and cytokinin of rocket plant (Erusa vesicaria). Khan et al. (2017b) suggested growth promoting potential of fresh and stored Moringa oleifera leaf extract in improving seedling vigor, growth and productivity of wheat crop. Yasmeen et al. (2012) reported performance of late sown wheat in response to foliar application of Moringa oleifera lam. Leaf extract. Rashid et al. (2018) studied foliar applied moringa leaf extract induces terminal heat tolerance in Quinoa. Combined application of moringa leaf extract and chemical growth - promoters enhances the plant growth and productivity of wheat crop (Triticum aestivum L.) suggested by Khan et al. (2020). Growth promoting potential of fresh and stored Moringa oleifera leaf extracts in improving seedling vigor, growth and productivity of wheat crop reported by Khan et al. (2017). Moringa leaf extract improves biochemical attributes, yield and grain quality of rice (Oryza sativa L.) under drought stress concluded by Khan et al.(2021).

Almehemdi et al.(2011) using licorice, fenugreek extracts and GA3 on yield of Caraway (Carum carvi L.). Effect of garlic and licorice extracts and urea on vegetative and flowering growth, yield and qualitative characters of cucumber (Hussein, 2002). Al- Sahaaf (2002) studied effect of licorice extract spraying on cracks types of pomegranate fruits. Al- Mersumy et al. (2001) reported effect of gibberellins, licorice extract and nutrients spraying on onion seed yield. Al- Jebouri et al. (2010) suggested role of spraying with some plant extracts in flowering of cucumber. A natural product containing fenugreek, tea and rosemary having fungus inhibiting and growth promoting for improving plant production and its use suggested by Hochenhull (2007).

Large distribution of W. arrhiza plants with its known detrimental importance is also thriving with great potential to multiply yield and alter quality of lentil crop. The conclusion are of paramount academic and applied significance, and are proven with promising prospects for utilization of W. arrhiza extracts by growers of commercial crop of lentil for higher and better quality lentil production.

Conflict of Interest

The author declare that he has no conflict of interest.

Author Contribution

This is the single author paper. All contribution are in this paper is Dr. Amit Tiwari

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Effect of Water extracts of W. arrhiza on vascular tissue in stem of lentil (Lens culinaries vr. D.P.L.,- 15)

Parameter	Level of Water extracts of W. Arrhiza
Parameter	Control	1%	2%	5%	C.D.
Average diameter of stem (µ)	1932.7	1947.1	1988.34	2012.4	1.8
Average thickness of stele (µ)	416.18	426.32	438.82	472.04	4.1
Number of Vascular bundles per-microscopic field	5	6	6	6	1
Average diameter of Xylem Vessels (µ)	28.42	28.88	31.56	29.94	1.8
Average diameter of Phloem (µ)	12.62	12.78	13.92	12.88	1.1

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Study of the Anatomical Growth Parameters of lentil Plant to Pre-soaked Seed treatment with Water Extract of Wolffia arrhiza

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