The effect of giving colchicine on appearance phnotypes of two genotypes in okra (Abelmochus esculentus L. Moench)

Cultivation of okra (Abelmochus esculentus L. Moench) is done to increase productivity by means of mutations to produce populations that have diversity. The aim of this study was to determine the effect of colchicine concentration on phenotypic changes in two genotypes of okra so as to produce plants with polyploidy properties. This research was conducted using a Randomized Block Design (RBD) with 2 treatment factors. First factor is use IPB’s Zahira Varieties: IPB’s Naila Varieties. Second factor was addition with a dose of colchicine 0%, 0.20%, 0.40%, and 0.60% as a solution for soaking okra seeds. Results showed that stomatal density (mm2) and flowering age WAP were significantly different due to variety and colchicine treatments. Variety significantly affected plant height at 5-13 WAP, number of leaves at 6-13 WAP, fruit length, fruit diameter, fruit weight, and number of fruit, colchicine concentration significantly affected height 2-13 WAP, number leaves at 4-9 WAP, flowering age, fruit length, fruit diameter, fruit weight, and number of fruit. Interaction of varieties and administration of colchicine concentrations affected okra plant height at 11-13 WAP, number of leaves at 8 WAP, and fruit weight. The qualitative characters show the results of changes in leaf color and shape.


Introduction
Okra (Abelmochus esculentus L. Moench) or better known as the name chickpeas, originally from West Africa, has even been around since Hundreds of years ago.This plant is spread to various tropical and subtropical regions Such as India, Japan, America, France, and Brazil, which in the end are more Popular in those countries [9] The main obstacle to okra cultivation in Indonesia is its unfamiliarity okra plant thoroughly by the people of Indonesia both benefits and Great potential to be developed because it has sufficient economic value High in horticulture.In addition, there is no variety yet Superior okra plants so that it can make it easier its processing [6].
Okra seed oil is rich in unsaturated fatty acids such as oleic acid and linoleic acid.Okra includes green vegetables that are rich in dietary fiber.In addition to fiber, Okra also contains 1.0 mg of glutathione and iron in 100 g of okra fruit young.Fiber is very important for the body because it can prevent constipation,obesity, hypercholesterolemia high cholesterol, diabetes and cancer [10].Okra (Abelmochus esculentus L. Moench) has been known as a plant multipurpose because almost all parts of the plant can be utilized [11].Mutation can be distinguished by cytological mutations, namely changes in shape, size, or number of chromosomes, and stomata as well as gene mutations that are cytologically absentappear but affect the phenotypic appearance.Colchicine (C22H25O6N) is a white alkaloid obtained from plants Cholchichun autumnale L (Family liliaceae) Colchicine (C22H25O6N) [8].Artificial mutations (induced mutations) occur as a result of human manipulation with chemical, physical and biological materials.Colchicin can block the formation of spindles cell division so that chromatid pairs in the metaphase phase of the process Cell division cannot separate and eventually polyploidy is produced [4].High productivity is certainly inseparable from the organs Plants are leaves.The main function of leaves is as a place of occurrence photosynthesis as well as exporting the results to all parts of the plant [5].
Soaking seeds in a solution of colchicin for a certain time (12-24 clock) will increase changes in the morphological shape of plant.Morphologically indicates a greater increase in plant height.It is indicates that the beam transporting xylem and phloem will enlarge as a result of enlarged cells [15].Stomata according to the function, shape, size and arrangement of epidermal cells It is not the same or different in different types of plants, nor is it the case stomata form or type.The aim of this research is to determine the effect of colchicine concentration [2].

Material and methods
This research was carried out in the research field of the Faculty of Agriculture, Universitas Sumatera Utara, Medan with an altitude of ± 32 meters above sea level.And carried out in the Faculty Biology Laboratory Mathematics and Natural Sciences, Universitas Sumatera Utara on altitude of place ±25 meters above sea level.Starts in the month March to June 2023.

Estimation of genetic parameter
The research data were analyzed using a Randomized Block Design (RBD) 2 factors based on the linear model as follows:

Correlation
Density calculation: stomata can be counted using the axiovision Rel 4.8 application.

Results and Discussions
The results obtained in (Table 4, 5, 7, 8, 9, 10) show that varietal factors can improve some intensity parameters because according to [3] states that each variety consists of a number of different genotypes and has different adaptability to certain environments.The results obtained in (Table 2, 4, 6, 7, 8, 9, 10) show that the factor in the form of colchicin concentration can increase several intensity parameters because this according to [1] states that colchicine as a mutagenic substance in polyploid will function to increase the size The results obtained in (Table 4.9) show that the 2 factors show a reciprocal interaction which can increase the 2 intensity parameters because in this study it was found that 0.60% colchicin administration was the best fruit weight in okra plants [7].One of the roles of potassium is to stimulate the translocation of assimilate from the source (leaves) to the storage organ (sink).[ 1 shows that the stomata density with the highest density was found in the V1K0 treatment with a density of 3,066 mm 2 which was significantly different from V2K3 with the lowest stomata density, namely 1,533 mm 2 .This is according to [16] which states that cells with higher ploidy are generally characterized by an increase in stomatal size Diameter of stomata.Table 2 shows that the stomata diameter with the highest mean was found in V2 treatment with an average of 1.59 mm and the lowest average was found in V1 treatment with an average of 1.56 mm.treatment accordance with the statement from [12] which states that the length of the stomata is related to the number of chloroplasts in the guard cells.Note.Numbers followed by the same letter in the same row and column are not significantly different according to DMRT at the α = 5% level.
Table 3 shows that okra plants aged 13 WAP in the Zahira (V1) variety treatment produced the highest plant height with an average of 58.25 cm, which was significantly different from the Naila (V2) variety treatment which produced the lowest average of 55.01 cm.The treatment of giving colchicine concentration showed that giving colchicine concentration at a dose of 0.60% (K3) resulted in the highest plant height.Table 4 shows that treatment varieties and colchicine concentrations and their interactions were not significantly different.Table 5 shows that the highest average flowering age was in the Naila (V2) treatment with an average of 54.29 DAP, which was significantly different from Zahira (V1) with an average of 50.62 DAP.In the colchicine concentration treatment, the highest average flowering age was in the K3 treatment with an average of 57.25 DAP, which was significantly different from the other treatments and the lowest average was in the K0 treatment average of 49.00 DAP.Note.Numbers followed by the same letter in the same row and column are not significantly different according to DMRT at the α = 5% level.
Table 6 shows that the fruit length with the highest average was in the Naila (V2) treatment with an average of 16.32 cm and the lowest average was in the Zahira (V1) treatment, namely 16.05 cm.Giving colchicine concentration to fruit length parameters in the K3 treatment had the highest average, namely 17.35 cm, which was significantly different from the K0 treatment which produced the lowest plant height, Table 7 shows that the fruit diameter with the highest average was in the Naila (V2) treatment with an average of 22.82 mm and the lowest average was in the Zahira (V1) treatment, namely 21.94 mm.Giving colchicine concentration to fruit diameter parameters in the K2 treatment had the highest average, namely 23.29 mm and the lowest average was in the K0 treatment, namely 20.68 mm.Note.Numbers followed by the same letter in the same row and column are not significantly different according to DMRT at the α = 5% level.
Table 8 shows that the weight of the fruit with the highest flattening.There is a V2 treatment with an average of 189.79 gr which is significantly different from V1 with the lowest average fruit weight of 158.9 gr.On concentration treatment Colchicin fruit weight with the highest average in K3 treatment with grade 248.03 gr which is significantly different from the K0 treatment with an average of 122.4 gr.The interaction of varieties in combination V2K3 treatment with flattening 189.79 gr.Table 9 shows that the number of fruits with the highest average was in the Naila (V2) treatment with an average of 17.75 fruits, which was significantly different from Zahira (V1) with the lowest average number of fruits, namely 16.83 fruits.Giving colchicine concentration to the number of fruit parameters in the K3 treatment had the highest mean, namely 21.16 pieces, which was significantly different from the other treatments and the lowest mean was in the K0 treatment, namely 13.83 pieces.

Qualitative Data
On qualitative characters show the results of changes in leaf color and leaf shape.This is according to [14] which states that another new plant formation from the appearance of phenotypes due to the influence of colchicin is the emergence of new form characters in plants.The research results showed that the colchicine concentration treatment causes changes in fruit color variations.The research results showed that the colchicine concentration treatment causes changes in shape or was the same as the control plants, namely round in shape.

Conclusions
Variety has a significant effect on density observation parameters stomata, plant height and number of leaves at 13 WAP.Giving colchicine concentration has a significant effect on diameter stomata, plant height at 13 WAP, flowering age, fruit length,fruit diameter, fruit weight, and number of fruits.The interaction between varieties and the concentration of colchicine has a significant effect on plant height and fruit weight.Plant height with the best combination is V1K3 and the best fruit weight combination is V2K3.The qualitative characters show the results of changes in leaf color and leaf shape.

Table 1 .
13]Stomata density of okra plants on varieties and colchicine concentrations.

Table 2 .
Stomata diameter of okra plants in varieties and concentration of colchicine.
Note.Numbers followed by the same letter in the same row and column are not significantly different according to DMRT at the α = 5% level.

Table 3 .
Okra plant height in varieties and concentration of colchicine.

Table 4 .
Average number of leaves (fruit) of okra plants for varieties and colchicine concentrations. .Numbers followed by the same letter in the same row and column are not significantly different according to DMRT at the α = 5% level. Note

Table 5 .
Age of flowering of okra plants in varieties and concentration of colchicine.
Note.Numbers followed by the same letter in the same row and column are not significantly different according to DMRT at the α = 5% level.

Table 6 .
The length of the fruit of the okra plant in varieties and the concentration of colchicine.

Table 7 .
The diameter of the fruit of the okra plant in the variety and the concentration of colchicin. .Numbers followed by the same letter in the same row and column are not significantly different according to DMRT at the α = 5% level. Note

Table 8 .
Fruit weight of okra plants on varieties and concentration of colchicine.

Table 9 .
Number of okra plant fruits in varieties and colchicine concentration.