The Effect of Spraying with Jasmonic Acid and Sugar Alcohols on some Chemical and Hormonal Properties of Lemon and Benzher Seedlings

In 2022, researchers from the University of Samarra’s Department of Horticulture and the College of Agriculture built a wooden canopy to perform their experiment. To study the effect of spraying with jasmonic acid and sugar alcohols on some chemical and hormonal properties of lemon and benzher seedlings, the seedlings were chosen as homogeneous as possible in size at the age of one and a half years. The experiment included three factors: the first being two Speciess of citrus seedlings, lemon and benzher. The second factor is spraying with three levels of jasmonic acid 0, 10, 20 mmol L-1, and the third factor is spraying seedlings with three concentrations of sugar alcohols 0, 3, 6 ml L-1. The experiment was designed according to a randomized complete block design (RCBD), the results showed the superiority of lime over benzher in the leaves content of nitrogen, chlorophyll, and jasmonic leaves content, and the values were 2.332%, 12.12 mg gm-1, 64.78 mg kg-1, respectively. As for benzher, it excelled in the rate of increase in the percentage of phosphorus, potassium, Carbohydrates, as it recorded 0.476%, 1.240%, and 3.284%, respectively.Also spraying with jasmonic resulted in a significant increase, especially the treatment Ja2, which was superior to the two treatments Ja1 and Ja0 in the leaves content of nitrogen, chlorophyll, leaf content of jasmonic, percentage of phosphorus, potassium, carbohydrates, and the results were 2.364%, 12.34 mg gm-1, 69.13 mg kg -1, 0496%, 1.263%, 3.300%, respectively. Spraying with sugar alcohols had a significant effect at the concentration of 6 ml L-1 S2 in the leaf content of nitrogen, chlorophyll, phosphorus, potassium, and carbohydrates, and the values were 2.339%, 12.20 mg gm-1., 0.491, 1.243%, 3.277%, respectively, while the average of these characteristics was lowest in the control group that received treatment S0. All of the qualities analyzed displayed substantial variations in the binary and triple interactions of the research components.


Introduction
Citrus trees are extensively distributed throughout the world, and they originated in the Malay Islands, as well as in parts of India and Vietnam, from where they later spread to other parts of the world [1].
IOP Publishing doi:10.1088/1755-1315/1259/1/012057 2 Most of the widely consumed fruits, including lemon, orange, bitter orange, lanky, sindhi, grapefruit, and other species, belong to the genus Citrus, which is regarded as one of the most significant genera in the Rutaceae family.One of the most significant fruit crops produced is citrus, which is grown extensively in more than 140 nations throughout the world in the tropical, subtropical, and subtropical areas, and between latitudes 40 north and south of the equator [2].The citrus lemon is a member of the genus Citrus, and its native habitats include southwest China and northeastern India.The third-most significant species of citrus is the lemon, a local variety that is popular in Iraq due to its tasty fruits, thin peel, and lower acidity than the other varieties [3].Citrus aurantifolia lemon is also referred to as Egyptian lime, major lime, West Indian lime, and Mexican lime.It is also known in Iraq as (Nomi Basra).When fully grown, the peel is thin and thin, and when it is ripe, it turns lemon yellow.The fruit's pulp is luscious and very acidic.China, Iran, India, Mexico, Brazil, Argentina, America, Spain, Italy, and Turkey are the primary countries where it is grown, those with an interest in plants concentrated on the commercial application of plant growth regulators, such as jasmonic acid, to enhance the vegetative development characteristics and chemical composition of the plant.As it activates the antioxidant systems responsible for resistance, stress can be resisted [4][5][6].One of the growth regulators and a naturally occurring substance in plants is jasmonic acid.It is regarded as a volatile organic compound that plays a significant role in regulating physiological processes and plant development in addition to its role in boosting the immune system against various pathogens and environmental stresses, which helped improve the vegetative growth of trees and the quality of fruits while lowering the risk of physiologic damage [7,8].Sugar alcohols are crucial natural nutrients for plants because they increase the time needed for micronutrients to be absorbed before drying, notably boron on the surface of the leaves.This improves foliar absorption.Spraying sugar alcohols with boron facilitates the process of transporting the element by forming a complex compound called Mannitol borate ester or Sorbitol borate ester at the sites of action of the element through the bark from the source to the downstream, increasing the vegetative growth and development of the yield and an increase in yield production [9,10].

Material and Methods
To determine the influence of spraying with jasmonic acid and sugar alcohols on some chemical and hormonal features of lemon and benzher seedlings, an experiment was conducted in the wooden canopy of the Department of Horticulture, College of Agriculture -University of Samarra during the season of 2022.A total of 108 seedlings of homogeneous vegetative growth were identified for experimentation , seedlings were age of one and a half years.andirrigated with a drip irrigation system.Also, all horticultural service operations were conducted on the seedlings until the end of the experiment.

The Field Experiment Included Adding seedlings Treatments
2.1.1.The First Being Two Speciess of Citrus Seedlings ( Lemon and Benzher) It has been symbolized (V 1 , V 2 ) in succession.

Jasmonic Acid
The leaves were sprayed with jasmonic acid at 0, 10, and 20 mmol.L -1 in three separate batches every 21 days beginning on 1/3/2021.The seedlings were sprayed thoroughly with water using a 10-liter holder sprayer first thing in the morning to increase the chances of the nutrient solutions sticking to the leaves and the seedlings living through the day.The effectiveness of the sprayed Jasmonic acid is improved by lowering the surface tension of the water particles.The code for the transactions was (Ja 0 , Ja 1 , and Ja 2 ) sequentially.

Sugar Alcohols
The sugar alcohols in the following concentrations (0, 3, 6 ml) contain organic sugar alcohols at 35.00% w/v, calcium (ca) water soluble at 2.10% w/v, and mannitol at 6.00% w/v.L -1 ) as in Jasmonic acid, the sugar alcohols treatments have been symbolically represented as (S0, S1, S2), along with the spreading substance used and the date of spraying.

Statistical Analysis
The trial followed a Randomized Complete Block Design (RCBD), with three treatments (two sets of three for a total of eighteen treatments).There were a total of 108 seedlings used in the study, with each pair of seedlings functioning as a separate experimental unit.The data was analyzed using an ANOVA table, with the components and their interactions tested in SAS, and the Least Significant Difference (LSD) calculated at the 0.05 level of significance [11,12].

Study Characteristics 2.3.1. Nitrogen Content in the Leaves %
I took 0.20 gm of the crushed sample was taken, to which 4 milliliters of sulfuric acid and 1 milliliter of pyrochloric acid were added, and the nitrogen was estimated using the (microcalculator) according to the method mentioned in [13].

Phosphorous Content in the Leaves %
The concentration of phosphorous was measured according to the method of [14], by taking 5 ml from the aforementioned digested sample beaker and treating it according to the spectrophotometric Vanadium Phosphomolybdate Method for measuring The visible density of phosphorus using a spectrophotometer (type -Bichrom 2005 Libra S22 -UK) at wavelength 420 nm, and using the standard phosphorus curve extracted the phosphorus concentration and then estimated the percentage.

Potassium Content in the Leaves %
Potassium was determined according to [15] method using a flame emission photometer.

Chlorophyll Content in the Leaves (mg g -1 )
The scientist's method [16] was used to estimate the amount of chlorophyll in the leaves.The leaves were washed well to remove dirt and dust from the surface of the paper, then 1g of fresh leaves were weighed and mashed by a clean ceramic mortar with (50 ml) 96% methanol, then the extract was separated by centrifugation at 1000 rpm and then filtered by Two layers of gauze were separated by centrifugation again at 2500 cycles/10 minutes, and the amount of absorption was measured by a spectrophotometer for measuring optical density (OD Optical Density) for chlorophyll A (663 nm) and chlorophyll B (645 nm), then the absorption density was calculated by applying the equation the following:-

Carbohydrate Content in the Leaves %
Carbohydrate content of the leaves was estimated according to the method of the scientist [14] By taking 100 mg dry weight of leaves for each treatment and placed in a water bath for three hours after adding 5 ml of 2.5 normal hydrochloric acid.Then, left to cool at room temperature and it was neutralized by solid sodium carbonate until the reaction stopped.The volume was completed with distilled water to 100 ml and discarded Centrifuge by centrifuge ( Hettich EBA 35, German origin ), 0.1 ml was taken from it and filled with distilled water to 1 ml, 1 ml of 5% phenol and 5 ml of sulfuric acid 96% were added to it.The solution was placed in a vibrator for 10 minutes after that in a 25-30 C water bath for 20 minutes and left to decrease in temperature and the appearance of orange indicates the positive reaction.Using the standard curve of glucose prepared previously, the percentage of carbohydrates was calculated using a spectrophotometer (Bichrom 2005 -Libra S22 -UK, British origin) at the light absorption 490 nm.

Jasmonic Acid Content in Leaves (mg.kg -1 )
This characteristic was measured with a Gas Chromatography-Mass spectrometer (GC-MS) according to the method of the scientist [17].

Nitrogen Content in the Leaves %
Table (1) displays the findings of a statistical analysis revealing that there were statistically significant variations between Species with respect to the percentage of nitrogen in the leaves of seedlings, with the best results being shown with the V 1 treatment, where the amount of nitrogen reached 2.332%.Treatment V 2 yielded the lowest amount, at 2.325%, whereas treatment Ja 2 , which was sprayed with jasmonic acid, resulted in a larger amount of nitrogen in the leaves, at 2.364%.Treatment Ja 1 comes in second with a value of 2.332%, very slightly higher than the 2.289% achieved by Comparison Treatment Ja 0 .Spraying with sugar alcohols also had positive differences as treatment S 2 achieved the best number of 2.339, followed by treatment S 1 which gave 2.331%, while Comparison Treatment S 0 achieved the lowest values for this trait, which amounted to 2.314%.Nitrogen content was discovered to be positively affected by the interplay of Species and Jasmonic spraying.The best results (2.369%) were obtained with the V 1 Ja 2 treatment, while the worst results (2.286%) were obtained with the V 2 Ja 0 treatment.Statistical analysis shows that spraying sugar alcohols on plants of different species led to a significant increase in leaf nitrogen, with treatment V 1 S 2 producing the highest value of 2.343% and treatment V 2 S 0 producing the lowest value of 2.309%.Nitrogen was also positively affected by the synergistic effects of Jasmonic acid and sugar alcohols spraying, with the highest value being 2.383% for the Ja 2 S 2 treatment and the lowest value being 2.275% for the Ja 0 S 0 treatment.As for the triple interaction between Species, Jasmonic acid and sugar alcohols, Results of Table (1) showed that the V 1 Ja 2 S 2 treatment was significantly superior by recording the highest number of 2.393% , while treatment V 2 Ja 0 S 0 achieved the lowest number of 2.270%.
Table 1.Effect of spraying with Jasmonic acid and sugar alcohols and the interaction between them on the nitrogen content in the leaves % of lemon and benzher.

Phosphorus Content in the Leaves %
Table (2) shows that compared to treatment V 1 , which resulted in a lower rate of phosphorus in the leaves (0.461%), treatment V 2 of Species was preferable.Treatment Ja 2 was the only one to increase the phosphorus content of the leaves to 0.496%, demonstrating that jasmonic acid spraying increased leaf phosphorus content.Then, treatment Ja 1 gave a value of 0.479%, compared to the treatment of control Ja 0 , which gave the lowest value of 0.429%; similarly, it was discovered through the same table that spraying with sugar alcohols significantly boosted the same trait, as treatment S 2 attained the highest value, reaching 0.491%, followed by treatment S 1 at giving it a value of 0.478%, compared to the treatment of control S 0 , which gave the lowest value of 0.429%.Phosphorus showed a positive difference in character when species interacted with jasmonic acid; the V 2 Ja 2 treatment had the highest value of 0.504%, while the V 1 Ja 0 treatment had the lowest value of 0.422%.Similarly, species interacted favorably with spraying sugar alcohols, with the V 2 S 2 treatment having the highest value of 0.504%.As for the triple interaction between Species , Jasmonic acid and sugar alcohols , results of Table (2) showed that the treatment V 2 Ja 2 S 2 was significantly superior by obtaining the highest number of 0.517%, while treatment V 1 Ja 0 S 0 achieved the lowest number of 0.340%.Table 2. Effect of spraying with Jasmonic acid and sugar alcohols and the interaction between them on the phosphorus content in the leaves % of lemon and benzher.The spraying of Jasmonic acid resulted in beneficial changes, with Treatment Ja 2 achieving the highest percentage of potassium content in the Leaves, at 1.263%, and Treatment Ja 1 achieving the lowest percentage, at 1.210%.The species had an apparent moral effect on the percentage of potassium content in the Leaves, as shown in Table (3), where Treatment V 2 recorded the highest percentage, at 1.240%, and Treatment V 1 gave the lowest percentage, at 1.210%.Table (3) shows the ethical effect of the interaction between Species and sugar alcohols, with treatment V 2 S 2 giving the highest percentage of 1.257% and treatment V 1 S 0 giving the lowest percentage of p.167%.We also note that the second interaction between the Species and spraying with Jasmonic acid had a significant effect, as the V 2 Ja 2 treatment gave the highest percentage of 1.270% and the treatment V 1 Ja 0 achieved the lowest percentage of p.1.167%.Treatment Ja 2 S 2 demonstrated a beneficial effect by providing the highest value, 1.275%, for the bilateral interaction between jasmonic acid spray and sugar alcohols, whereas Treatment Ja 0 S 0 provided the lowest value, 1.135%.

( V ) Species
Table 3 shows that the species, jasmonic acid spraying, and sugar alcohol spraying all worked together to significantly boost the proportion of potassium content in the Leaves, with the V 2 Ja 2 S 2 treatment producing the highest percentage (1.280%) and the V1Ja0S0 treatment producing the lowest percentage (1.180%).Table 3.Effect of spraying with Jasmonic acid and sugar alcohols and the interaction between them on the potassium content in the leaves % of lemon and benzher.Based on the data in Table (4), we can see that the application of jasmonic acid spray had a beneficial effect and led to an increase in the chlorophyll content of the leaves (Treatment Ja 2 achieved the highest conc. of chlorophyll per leaf, at 12.12 mg g -1 ), while the application of the species (Treatment V1) had no effect (Treatment V2 had the lowest conc. of chlorophyll per leaf, at 12.06 mg.It was also discovered that spraying with sugar alcohols significantly affected the chlorophyll content of the leaves, with treatment S 2 producing the highest amount of chlorophyll (at 12.20 mg g -1 ), treatment S1 producing 12.08 mg g -1 , and treatment S 0 producing the lowest amount (at 11.99 mg.g -1 ).Table (4) shows that there is a statistically significant difference in the chlorophyll content of leaves due to the interaction effect between species and jasmonic acid spraying, with the V 1 Ja 2 treatment producing the highest rate of 12.47 mg g -1 and the V 1 Ja 0 treatment producing the lowest rate of 11.77 mg g -1 .The maximum overall Chlorophyll content of 12.23 mg g -1 was found in the V 1 S 2 treatment, indicating a clear substantial increase due to the bilateral interaction between the species and sugar alcohols (see the same table for details).The Chlorophyll concentration of V 1 S 0 was 11.97 mg g -1 , the lowest of any treatment.The table also reveals a noteworthy impact due to the bilateral interaction between jasmonic acid and sugar alcohols, with treatment Ja 2 S 2 producing the highest Chlorophyll content of 12.50 mg g -1 and treatment Ja0S0 producing the lowest at 11.70 mg g -1 .

( V ) Species
Table (4) shows that there are substantial variations in leaf Chlorophyll content due to the triple interaction, with the V 1 Ja 2 S 2 treatment yielding the greatest Chlorophyll content of 12.60 mg g -1 .
However, the Chlorophyll content was 11.60 mg g -1 lowest for the treatment V1Ja0S0.