Diversity induction with gamma-ray irradiation on Phalaenopsis amboinensis var. Common natural orchid generation M1V0

Phalaenopsis is a popular genus in the Orchidaceae family. Phalaenopsis amboinensis orchid is an endangered natural orchid, so it needs to be preserved. Conservation of natural orchid Phalaenopsis amboinensis var. Common way is by increasing genetic diversity. This research aims to determine the characteristics and morphological diversity of Phalaenopsis amboinensis natural orchid irradiated gamma-ray results and without gamma-ray irradiation; this research was conducted in two places: the implementation of gamma-ray irradiation at the PAIR-BATAN Laboratory and in Matesih, Karanganyar with an average height of 450 m above sea level. The research was conducted from December 2022 to May 2023. The study used a simple design of one factor, the doses of gamma-ray irradiation with 5 levels: 0 Gy (R0), 7.5 Gy (R1), 15 Gy (R2), 22.5 Gy (R3), 30 Gy (R4). Each treatment contained five accessions. So, there are 25 experimental units. Observational data were analyzed descriptively by comparing observational data from Phalaenopsis amboinensis var. Common Generation M1V0 orchids irradiated with control plants. The results showed diversity in the morphology of the Phalaenopsis amboinensis var. Common orchid Generation M1V0 results from gamma-ray irradiation based on qualitative morphology with phenotypic observations, showing two orchid leaf shapes: oblanceolate and elliptic. There are accessions with fasciated root conditions.


Introduction
The diversity of orchids in Indonesia is very high, and they are ornamental plants with high economic value and a variety of flower shapes and colors [1].One of the orchids of the Orchidaceae genus is Phalaenopsis.Phalaenopsis orchids are in great demand because of their attractive colors, flowers that can bloom for a long period, and various shapes and color patterns; they often flower because the flower stalks, after flowering, can grow new stalk with flower buds or even new plant buds called Keiki [2].The existence of Phalaenopsis orchid species or forest orchids is decreasing, so it is necessary to preserve them [3].The Phalaenopsis amboinensis orchid is a species of moth orchid that is endangered.Phalaenopsis amboinensis is an epiphytic orchid with a monopodial growth type originating from tropical climates at an altitude of 500-700 m.Phalaenopsis amboinensis from Sulawesi has yellow IOP Publishing doi:10.1088/1755-1315/1317/1/012001 2 flowers with fine brown lines, while white flowers with thick brown or reddish brown lines are found in Maluku with the variant name Common [4].Preserve the Phalaenopsis amboinensis orchid through gamma-ray irradiation.Gamma radiation mutagenesis is one way to obtain new genetic diversity from plant species [5].Several research results show that gamma-ray irradiation can change the morphological characteristics of plants.Research with irradiation treatment with several doses of gamma rays must be carried out on Phalaenopsis amboinensis var.Common Generation of M1V0 to produce morphological diversity [6].

Materials And Method
The natural orchid used is the orchid Phalaenopsis amboinensis var.Common, which comes from Maluku.The research was carried out in two places the irradiation was carried to the implementation of gamma-ray irradiation at the PAIR-BATAN Laboratory, Jakartaa Selatan, DKI Jakarta and Acclimatization in Matesih, Karanganyar, with an average height of 450 m above sea level [7].Growing Phalaenopsis plant optimally at a temperature of 24 o C-28 o C. Temperature below 25 o C [8].The research was carried out from December 2022 to March 2023.The Gamma Ray radiation dose treatment consisted of 5 levels: 0 Gy (R0), 7.5 Gy (R1), 15 Gy (R2), 22.5 Gy (R3), 30 Gy (R4).Each treatment contained five accessions, so there were 25 experimental units.The data obtained was then analyzed.The qualitative method was carried out descriptively by observing the phenotype of each orchid plant in various doses of Gamma-ray irradiation and controls.In contrast, the quantitative method used box plots to see diversity.The moth orchid is known from its morphology as a characteristic [9].

Qualitative Morphological Diversity
The research obtained qualitative data using phenotypic observations per the morphological characterization descriptors of orchid plants.Morphological characterization is carried out on the appearance of plants, such as leaf characters starting from the leaf tip, leaf edge, leaf shape, and leaf texture [10].Quantitative data is based on size characters from morphology, and qualitative data is based on morphological characters according to the descriptor table.Phenotypic observations were made on leaf shape, cross-section, leaf tip, leaf arrangement, leaf edge, leaf surface texture, and leaf symmetry in all accessions.This phenotype observation data was carried out at week 12 since the growth of new leaves after irradiation.
The leaf shape of the orchid Phalaenopsis amboinensis var.Common Generation M1V0, according to the results of phenotypic observations, shows diversity in leaf shape; the accessions show two shape types: oblanceolate and elliptic.The oblanceolate shape was in all accessions with irradiation doses 0 Gy, 7.5 Gy, and 22.5 Gy.The treatment dose of 15 Gy showed that the leaves of accession A1, A2, and A5 were elliptic in shape.The elliptic leaf form was also found in all the accessions with an irradiation dose of 30 Gy.The oblanceolate leaves are 6-12 cm long and 2-4 cm wide [11].Cross sections show the results of suppressed symmetry phenotypes in all accessions.Leaf tips of Phalaenopsis amboinensis var.Common Generation M1V0 in all accessions shows that the leaf tip is blunt.The leaf arrangement or phyllotaxis in all accessions shows a double type.Phalaenopsis orchids have leaves with the growth position of the leaves mounted and lined up in two close rows facing each other 180 o [12].The edges of the leaves on all asses show that the edges are flat.The surface texture of the leaves, according to the results of phenotypic observations, showed that all the accessions had bare leaf surfaces.In contrast, leaf symmetry showed that all the accessions had symmetrical leaves.The fasciation process can be seen in Figure 1, representing accession 1 treated with an irradiation on 7.5 Gy (R1A1).Figure 1 shows that the roots of the Phalaenopsis amboinensis orchid var.Common generation M1V0 shows thickening and flattening, then splitting.Unbalanced cell proliferation causes plant organs to fasciate, which reflects abnormal meristem proliferation, derived from the Latin word 'fascis', which means 'bundle' [13].Gamma irradiation of plants causes damage to plant tissue in the growing roots, stems, and shoots, resulting in morphological deviations that allow better characters to be obtained compared to their parents [14] and the formation of Phalaenopsis amboinensis var root organs.Common generation M1V0 begins to appear two months after irradiation, so Phalaenopsis amboinensis var.Common generation M1V0 has been mutated.In addition to the nucleus, plant cells also have gene expression systems in mitochondria and plastids.Gene expression in organelles is usually associated with organelle-specific functions, but also involves higher-order physiological activities such as the control of organogenesis [15].Plant organogenesis forms plant organs, such as roots, flowers, stems, and other organs.Fasciation is caused by extra cell division at an early stage and can occur in any part of the plant organ.Bacteria can cause fasciation and can also be caused by genetic disorders [16].

Leaf Number
The total leaf number was calculated from the growth of new leaves until the end of observation at week 12.The strength of the leaves to survive is the availability of nitrogen and water so they do not easily fall prematurely.The availability of absorbable nitrogen can influence the growth and number of leaves [17].The highest number of leaves in the 0 Gy treatment was 6 leaves, and the lowest was 3 leaves.In the 0 Gy treatment, the number of leaves ranged from 3-6 leaves.Irradiation by 7.5 Gy dose treatment showed that the lowest number of leaves was 4 pieces, and the highest number of leaves was 6 pieces.The 7.5Gy treatment had a leaf number range of 4-6 leaves.The 15 Gy treatment showed that the lowest number of leaves was 4 pieces, and the highest leaf number was 6 leaves.The 15 Gy treatment had a leaf number range of 4-6 leaves.The 22.5 Gy treatment showed that the lowest number of leaves was 3 pieces, and the highest was 6 pieces.The 22.5 Gy treatment had a leaf number range of 3-6 leaves.The 30Gy treatment showed that the lowest number of leaves was 4 pieces, and the highest was 5 pieces.The 30Gy treatment had a leaf number range of 4-5 leaves.The amount of nitrogen absorbed and carbohydrates formed so that the protoplasm, which becomes new cells, will increase [18].with gamma-ray irradiation The number of leaves for each accession.The range of boxplot bars determines the area of data distribution; the longer the data range, the wider the data distribution.The width of the data distribution shows diversity, so the longer the span of the boxplot bar shows that the resulting diversity is also higher.Figure 2 shows that irradiation with doses of 0 Gy and 22.5 Gy has a comprehensive data distribution, so the resulting diversity is high.The number of green leaves is calculated by the number of still-fresh leaves, not yellow or senescent.Leaf turgor affects the aging or senescence process; if leaves experience transpiration but do not receive adequate water and nutrients, this will speed up the senescence process [19].

Leaf Length
Leaf length is measured on newly growing leaves.The highest leaf length in the 0Gy treatment was 13cm, and the lowest was 12cm.In the 0Gy treatment, the leaf length range was 12-13cm.The 7.5 Gy treatment showed that the length of the lowest leaf was 10 cm and that of the highest leaf was 13.5 cm.The 7.5Gy treatment had a leaf length range of 10-13.5 cm.The 15Gy, 22.5Gy, and 30Gy treatments showed that new leaves did not grow, so no leaf length data was obtained.The leaf length data with the longest boxplot stem span is at a dose of 7.5 Gy, the leaf length of the orchid Phalaenopsis amboinensis var (Figure 3).Common M1V0 generations with various lengths are treated with an irradiation dose of 7.5 Gy.The box plot stem with the shortest span is in the 0 Gy treatment with leaf lengths that tend to be similar.Irradiation doses of 15 Gy, 22.5 Gy, and 30 Gy did not show any new leaf growth after irradiation, so no data on leaf length was obtained.Induced gammaray irradiation can affect orchids' physiological and morphological processes [20].The standard length of natural orchid leaves is 25cm to 40cm, according to the character of each Phalaenopsis orchid [21].

Leaf Width
Leaf width is measured on newly growing leaves.The highest leaf width in the 0Gy treatment was 3.5cm, and the lowest was 3.3cm.In the 0Gy treatment, the leaf length range was 3.3-3.5cm.The 7.5 Gy treatment showed that the length of the lowest leaf was 3.5 cm and that of the highest leaf was 3.7 cm.The 7.5Gy treatment had a leaf length range of 3.5-3.7 cm.The 15Gy, 22.5Gy, and 30Gy treatments showed that new leaves did not grow, so no leaf width data was obtained.The efficiency of leaf photosynthesis can produce morphological changes such as different leaf widths and lengths [22].The width of the leaves of the orchid Phalaenopsis amboinensis var.Common Generation M1V0 showed that accessions without treatment and those treated with a dose of 7.5 Gy showed the highest width at a dose of 7.5 Gy, reaching 3.7cm (Figure 4).The lowest width was shown in accessions without treatment, namely only 3.3 cm.Irradiation doses of 15 Gy, 22.5 Gy, and 30 Gy did not produce leaf width data.Mutations can cause polyploidy, resulting in morphological character changes [23].
Irradiation doses of 15 Gy, 22.5 Gy, and 30 Gy did not provide data on leaf width because what was calculated were new leaves that grew after irradiation.At doses above 7.5 Gy, growth is seen to be inhibited.Stunted growth can be due to unstable or disturbed physiological processes in plants.Irradiation is a solution to provide new genetic variations.These new genetic variations also affect physiological processes due to cell damage [24].

Conclusion
There is diversity in the morphology of the Phalaenopsis amboinensis var.Common orchid Generation M1V0 results from gamma-ray irradiation based on qualitative morphology with phenotypic observations, showing two orchid leaf shapes: oblanceolate and elliptic.There are accessions with fasciated root conditions.

3. 2 .
Another Conditions Phalaenopsis amboinensis var.Common Orchid Generation M1V0 indicates other conditions beyond the expected observed variables.The condition shows changes in root shape.Roots show fasciation conditions.Fasciation on the roots of the Phalaenopsis amboinensis orchid var.Common Generation M1V0 initially looks thick, then becomes wrinkled and flattened, then divides dichotomously.

Figure 2 .
Figure 2. Boxplot of Phalaenopsis amboinensis var.Common orchid generation M1V0 leaf numberwith gamma-ray irradiation The number of leaves for each accession.The range of boxplot bars determines the area of data distribution; the longer the data range, the wider the data distribution.The width of the data distribution shows diversity, so the longer the span of the boxplot bar shows that the resulting diversity is also higher.Figure2shows that irradiation with doses of 0 Gy and 22.5 Gy has a comprehensive data distribution, so the resulting diversity is high.The number of green leaves is calculated by the number of still-fresh leaves, not yellow or senescent.Leaf turgor affects the aging or senescence process; if leaves experience transpiration but do not receive adequate water and nutrients, this will speed up the senescence process[19].

Figure 4 .
Figure 4.The width of the leaves of Phalaenopsis amboinensis var.Common Generation M1V0 gamma-ray irradiation

Table 1 .
Qualitative morphological diversity of Phalaenopsis amboinensis var.Common Orchid

Table 2 .
Highest and lowest values on quantitative variables of Phalaenopsis amboinensis var.