Tissue Propagation of Marjoram Plant (in vitro)

The experiment was conducted in the plant tissue culture laboratory of the Department of Horticulture and Landscape Engineering, College of Agriculture, University of Kirkuk for the season 2022. to study the propagation of the marjoram plant tissue from tips of branches outside the body. plant parts were sterilized for a period of 10-15 min, water was used to remove dust, and they were sterilized inside the cultivation table in Naocl sodium hypochlorite solution (commercial minor solution) at a concentration of 6% for 8 minutes in order to remove suspended microscopic parts. plant parts were washed 3 times each for 5 minutes. plant parts were cut, and tips of branches were taken, with (1) cm. with kinetin at, 2, 4, 6, 8) mg. L-1. plants were vaccinated at a temperature of 25 + 2 m 0 with an intensity of illumination was 3000 lux, with a light sequence of 16 hours, followed by 8 hours of darkness, for 4 weeks of cultivation. data were taken for the experiment (length of the longest branch) (number of branches) (number of leaves). after replanting the tips of the branches for another 4 weeks of the multiplication process, data were taken at (length of the longest branch cm) number of leaves/plant part). tips of branches. Regarding the root experience, data were taken (percentage of rooting %) (average number of roots/vegetative part) (average root length/cm) (branch length/cm) (number of leaves/vegetative part) full and half concentration of salt from the same soil were added to cultivated on a medium with different concentrations of IBA.


Introduction
Marjoram, Origanum majoraua, is a perennial and aromatic herbaceous plant belonging to the Lamiaceae family.It is native to the Mediterranean is a native region [1].Marjoram has many health properties, including anti-inflammatory, antimicrobial, and analgesic.It has many active compounds, vitamins, flavonoids, glycosides, and Hydroquinone in addition to the triterpenes [2].Tissue culture is one of the biotechnologies that plays an important role in human service, especially in producing huge numbers of plants that's a free from pathogens and similar at one time.Relatively short and at any time, also used in research technology, including breeding and improvement plants, medicinal drugs, and rapid propagation of strains, and important to carry out by following different methods of differentiation and morphological formation, such as the formation of adventitious buds, stimulation of growth of axillary buds, development of asexual embryos (somatic embryos).study of basic aspects of plant growth development and secondary metabolism, [3][4][5][6].Tissue culture techniques (culture outside 1259 (2023) 012043 IOP Publishing doi:10.1088/1755-1315/1259/1/012043 2 the living body) during recent decades have spread widely in the world.It is obtained from plant extracts after increasing their quantity or concentration [7].

Materials and Methods
The experiment was carried out in the tissue culture laboratory/ Department of Horticulture and Landscape/ College of Agriculture/ University of Kirkuk.for season 2022 to study the propagation of the marjoram plant tissue (outside the living body).Leaves were taken and placed under water for 10-20 minutes in order to get rid of harmful pollutants and microscopic microorganisms.they were sterilized by the planting table in sodium hypochlorite (Naocl commercial solution) at a concentration of 6% for 8 minutes to remove suspended microorganisms.parts were washed 3 times with distilled water for 5 minutes each time to remove sterilizers from parts of plants.plant parts are cut from branch tips, each part has a length o 1 cm length, and grown on Ms medium supplemented with 3% sucrose and 6-7 g.Acars with (0, 2, 4, 6) mg.L -1 of kinetin.The plants were incubated at 2-25 m 0 and a light intensity of 3000 lux, with a light sequence of 16 hours, followed by 8 hours of darkness.after 4 weeks of cultivation, the data were taken for the experiment of emergence (length of the longest branch in cm) (number of branches) (number of leaves in the plant part) and after re-cultivation and 4 weeks later of the multiplication process, the following data were taken (length of the longest branch in cm) ( Number of branches ((number of leaves of the vegetative part) As for the rooting experiment, the following data were taken and after another 4 weeks of cultivation (percentage of rooting %)(the rate of roots (the roots of the vegetative part)) (the average length of the root cm) (the number of branches the vegetative part)( number of leaves of the plant part).results were analyzed according to the complete random design (CRD) and one treatment consisted of 10 replicates, one replicate contained one plant part the statistical analysis program used [7], to analyze the data compared using Duncan's multiple tests and under the probability level of 5% (Dun cans multiple) [8].Treatments and follow:

The Effect of NAOCL Concentrations on the Process of Evaluating Plant Parts
From the data in Table (1), it is clear that the duration of sterilization had an effect on the percentage of contamination, where the lowest percentage of pollution was 5% at a concentration of 6%(Naocl) after 8 minutes, while the highest was 50% at 2% after 15 minutes, as another percentage, the concentration 4% during 12 minutes, the survival was 70% and the contamination was 30%, based on the data in the The significant increase in the number of (uncontaminated) plant parts that are free from pollution is caused as a result of immersing them in the commercial bleach solution in Table (1).the reason may be attributed to the effectiveness of commercial bleach in sterilizing the plant, also the commercial minor material is characterized by its ability to be easily removed from parts of the plant By washing, being soluble into less toxic compounds, than easier to remove than the vegetable parts of the compound itself [7].These results are consistent with the findings of [8], when propagating the herbal rose plant, which when immersed in a diluted commercial minor solution helped reduce the percentage of plant contamination the researcher was able to obtain the highest percentage of healthy plant parts 92% for 10 minutes using Commercial binder at a concentration of 10%.These results agreed with many studies on the effectiveness of the commercial binder solution in sterilizing starters grown outside a living body.These experiments confirmed effectiveness of solution in obtaining healthy seedlings, [9].These results also agreed with what [10], reached when using a commercial minor in sterilization experiments.

The Effect of Kinetin Concentrations on the Development of the Tips of the Branches
The data in table (2) showed the effect of using kinetin on tips of branches growth, kinetin 6 mg.L -1 recorded a value of 2.7500 compared to the control treatment.kinetin 2 and 4 mg.L -1 gave 0.625, 0.875, 2500, in a number of branches, respectively.there are no significant between the last two treatments.All treatments were superior to the comparison treatment, although there were no significant differences Among the concentrations used, the longest branch length at the concentration 8 mg.L -1 , gave (1.875, 1.837, 1.2250), respectively.The used quinine had a significant effect on the number of leaves, as it recorded significant differences between the concentrations used and the comparison treatment, in addition to significant differences between the same treatments, where the treatment 8 mg.L -1 was superior to the rest and gave 20 leaves.(6.4) mg.L -1 recorded 13.500, 10.875, respectively.the concentration of 2 mg.L -1 recorded 3.500, 1.250, respectively.Table 2. Shows the effect of using kinetin on the formation of the tips of the branches.Means followed by the same letter (s) within each column during each season are not signifificantly different at 0.05.

Effect of Using Kinetin on the Doubling of the Tips of the Branches
The data in table (3) showed the effect of using Kinetin on the media to develop the tips of the branches, where the highest value was recorded at the concentration 8 mg.L -1 reached 3 mg.L-1 and superior all the treatments.followed by 2 mg.L -1 .while the control recorded the lowest value.while the concentration of 8 mg.L -1 did not exceed each of the concentrations (4,6) mg.L -1 and gave 2.2500.
2.21250 number of branches .As for the effect of Kin on the length of the longest branches, the concentrations (8, 6, 4, 2) mg.L -1 gave the highest value at the concentration 1.5125, followed by (6, 4, 2) mg.L -1 , a concentration which recorded 1.3625, 1.4375, 1.2000, respectively, while the control gave the lowest value of 1.1625.kinetin effect on a number of leaves, at a concentration of 8 mg.L -1 , recorded a significant difference compared to the control, followed by 2 mg.L -1 reached 11.625 leaves, which gave 2.250, 5.375.while there was no significant recorded between 8 mg.L -1 and (4, 6) mg.L -1 , concentration, when the number of leaves was (8.875, 7.750).) Respectively.
Table 3. Shows the effect of using kinetin on the doubling of the tips of the branches.Means followed by the same letter (s) within each column during each season are not signifificantly different at 0.05.
Cytokinins have a role to increase cell division, cells are unspecialized and undifferentiated in an initial developmental stage.a number of branches were obtained as a result of a state of hormonal balance between endogenous hormones and growth regulators added to the nutritional medium [11,12].Kinetin has a role in regulating enzymes that reduce nitrates and transport sugars, in addition, it works to increase number of leaves, branches, and dry weight of shoot [13].These results agreed with [8] by obtaining significant results in a number of branches, leaves, and plant height with high concentrations of Kinetin 3.4.Rooting of the resulting plants on MS medium half the concentration of salts and forged with IBA data table (4) showed the effect of using IBA on plant parts grown tissue in artificial culture media and a significant effect on the rooting percentage characteristic, where the highest rooting percentage was at the concentration 0.25 mg.L -1 and reached 80%, then the rooting percentage was 40% for each of the concentration (0, 5, 1) mg.L -1 , and these concentrations were superior to both the control treatment and the concentration of 2 mg.L -1 , which were recorded (0.30%), respectively, as a percentage of rooting.As for the number of roots.The treatment with 0.25 of IBA was superior to all treatments and gave the highest value of 5.375 number of roots, followed by 2.125, 1.125, 1.500, and as a followed by (0.5, 1, 2) mg.L -1 , while the control gave lowest number of roots at 5%.. IBA and auxin showed significant differences between the treatments and control, while no significant differences were recorded between the concentrations of IBA.The highest root length was recorded at the concentration of 0.25 mg.L -1 which reached 1.787.the concentrations of (0.5, 1, 2) mg.L -1 .were recorded at 1.250, 1.0375, and 1.3750 in root lengths, respectively.the highest value was recorded at the concentration of 0.25 mg.L -1 compared to the control treatment, was recorded at 12,500 in leaf number, followed by 10,250 and 11,250 at the concentrations (0, 1, 5) mg.L -1 As for the first number of leaves, it was recorded at a concentration of 2 mg.L -1 .Also, no significant effect was obtained as a result of using IBA in the branch length.The highest value of branch length was 2.862 at a concentration of 0.25, then followed by the concentration 0.5 mg.L -1 for comparison, which gave (2.255, 2.2500), while the lowest was recorded.At a concentration of 2 mg.L -1 , it was 2.212 cm.Means followed by the same letter (s) within each column during each season are not signifificantly different at 0.05.

Rooting of the Resulting Plants on MS Medium Complete the Concentration of Salts and Forged with IBA
It was clear from the data table (5) there was a different significance as a result of using the full concentration of salts and different concentrations of IBA, where all concentrations used in the rooting percentage were superior to the control, which gave 5%, while the highest percentage of rooting was at concentration 0.25 reached 50%, followed by the concentration (0.5, 1, and 2) mg.L -1 , which reached (40%, 30%, and 30%), respectively.The number of roots did not record any significant difference between the concentrations, compared to the control treatment, which gave 5% of number of roots, the largest number of roots was recorded at the concentration of 0.25 mg.L -1 , while the concentrations (0.5, 1, 2) mg.L -1 showed (1.125, 1.250, 0.875) root length.As previously, the different concentrations of BA did not record any significance between treatments, while the control was given a percentage of 5%.mg.L -1 (1.012, 0.787, 0.687) for the longest roots.BA had a significant effect on the number of leaves, use of a full concentration of salts, and treatment of 0.5 mg.L -1 was superior to the concentrations used for BA recording the highest number of leaves (12.500, 12.00), respectively.the lowest value was recorded at a concentration of 1 mg.L -1 at 8.500, then 9.250, 9.750, 0.5 at a concentration of 0.5, 2 mg.L -1 .The IBA had a significant effect on the branch length, where the treatment was 0.5 mg.L -1 was superior to all treatments compared to the control, the highest was recorded and reached 2.837 cm, while the lowest value was recorded at concentrations (0.25, 2) mg.L -1 2.112 Then the concentration was followed by 1 mg.L -1 compared to control, which gave (2.337, 2.262) cm.Table 5.Effect of total salt concentration of MS medium supplemented with IBA on rooting of plant parts.Means followed by the same letter (s) within each column during each season are not signifificantly different at 0.05.Indol Butyric acid (IBA) is a plant growth regulator that plays a role in the rooting process because of its prevalence in the plant.by the process of its transfer in plants [14], as well as the efficiency of IBA in the rooting process due to its high stability, the effect of growth [15,16] results are consistent with mention that use of [17] a salt concentration to increase number and length of apple roots.Agreed with [18] IBA with a half salt concentration from Ms medium was superior in increasing rooting rate of Bougainvillea glabra.

Conclusion
We concluded from this study that the highest concentration of 6% of NaoCl with the least time of 8 minutes is optimal for sterilization of explant, while the optimal concentration for the emergence and multiplication of explant is 8 mg.L -1 of Kinetin and includes the characteristics of the number of branches, number of leaves and the length of branches.While half of the concentration of salts from the MS medium with IBA was the best in the Root%, and at a concentration of 2 mg.L -1 of IBA, while the optimal concentration to obtain the longest root length and number was at a concentration of 0.25 mg.L -1 of IBA.

Table 4 .
Effect of the concentration of MS medium supplemented with IBA on the rooting of plant parts.

Table 1 .
table, then sterilization in the subsequent experiments Effect of using NAOCL on sterilizing plant parts.