EFFECT OF BIO HEALTH, TECAMIN MAX AND BENZYL ADENINE ON GROWTH CHARACTERISTICS OF SWEET ORANGE SEEDLINGS

This investigation was aimed to study the effect of soil application of Bio health and foliar application with Tecamin max and Benzyl adenine on growth characteristic of sweet orange ( Citrus sinensis L.) seedlings during two successive seasons (2020 and 2021). Bio health was added with three concentrations (0, 6 and 12 g.L -1 ) to the soil and foliar application of Tecamin max with three concentration (0, 5 and 10 ml.L -1 ), Benzyl adenine with three concentration (0, 100 and 200 mg. L -1 ) on the sweet orange seedlings which were brought from private nursery in Duhok city and have two-years old and nearly uniform in growth vigor. The results proved that all parameters such as increases in plant high, stem diameter, branch numbers, single leaf area, total chlorophyll as soon as leaves carbohydrate and leaves dry weight in both seasons were increased significantly as compared with control, accept Tecamin max had no significant effect on leaves carbohydrates and leaves dry weight, Benzyl adenine no significant effect on leaves dry weight on both seasons, Furthermore, combination among high concentration of Bio health, Tecamin max and Benzyl adenine improved all parameters in comparison with the control.


INTRODUCTION
Citrus fruits are one of the most delicious fruits belonging to the family Rutaceae.All commercially important species belong to the genus citrus including sweet orange, Mandarin, lime, Lemon, Sweet lime, Grape fruit, etc. Sweet orange is originated from southern China where it has been cultivated for many years (30).But today it is grown commercially worldwide in tropical, subtropical and some temperate regions to become the most widely planted fruit tree in the world (21).Nutrition is an important input that determines the soil health and production as a whole, the application of nutrients to both young and maturing trees determines the vegetative development of citrus trees (27).Today, chemical fertilizers are undesirable in crops nutrition because it had adversely effects on the soil fertility, water quality, yield and quality of the products (6,33).Therefore, uses of bio-fertilizers, amino acids and Benzyl adenine have assumed great importance for sustainable production and to improve the soil physical, chemical and biological properties.Also, they are a good alternative to reduce uses of chemical fertilizers.In this respect, several workers reviewed the significant role of bio-fertilizers in influencing the soil properties and enhancing the growth, yield and quality of citrus (17,24,25).Bio-fertilizers have the potential to increase the health and productivity of plant life and reduce the need to use synthetic fertilizers (7,8).Most of biofertilizers consist of microbes that are involved in the decomposition of organic matter and the breakdown of minerals into a soluble form that is useful to plants.The results Al-Abbasi.(1) showed that bio-fertilizers treatment of Azospirillium and Bacillius with organic fertilization was significantly increased the height of seedling, stem diameter, number of branches, number of leaves, leaf area and shoot fresh and dry weights compared with control treatment which gave the lowest values.
Amino acids are organic compounds; the key elements of an amino acid (C, (H, O and N), although other elements are found in the side chains of certain amino acids.About 500 naturally occurring amino acids are known (though only 20 appear in the genetic code) and can be classified in many ways (28).
Amino acids have various prominent functions in plants.Besides their usage during protein biosynthesis, they also represent building blocks for several other biosynthesis pathways and play pivotal roles during signaling processes as well as in plant stress response.In general, pool sizes of the 20 amino acids differ strongly and change dynamically depending on the developmental and physiological state of the plant cell (19,31).The results Al-Himdany.(3) showed that spraying of saplings with Disper chlorophyll had high significant increase in plant length -stem diameter, branch umber, leaves area, root number, root length, root branches compared to the control treatment.Benzyl adenine (BA) can be considered to be an endogenous-like compound which provides very little risk to the environment.A precondition of precocious bearing in young trees is the development of a canopy structure which has good cropping potential and this can be achieved by using BA.The efficiency of PGRs is determined by the physiological age of trees, by the environmental conditions at application and by the application methods used (12).Al-Juboori.
(5) investigated the effect of three concentrations of Benzyl adenine (0, 100, 200) mg.L -1 and three concentrations of Gibberllic acid (0, 150, 300) mg.L -1 on Two citrus seedlings (Swingle citrumelo and Troyer citrange).The results showed superiority of Swingle citrumelo root stocks in C/N ratio, phosphorus and potassium leaves content for two seasons while Troyer citrange root stock showed superiority in leaves content of (Nitrogen and Protein) which.the treatment of Benzyl adenine 200 ml.L -1 had significant increases in all studied characteristics C/N ratio, Nitrogen, Protein, phosphorus and potassium for two seasons (10.01, 1.533%, 9.57% ,0.247% and 0.602%) for season 2013(11.22,1.703%, 10.64%, 0.275% and 0.669%) for season 2014 respectively.This study was aimed to investigate the effect of Bio health, Tecamin max and Benzyl adenine alone or in combination on vegetative growth characteristics of sweet orange seedlings.

MATERIALS AND METHODS
This study was carried out in the lath house of the College of Agricultural Engineering Sciences, Duhok Province, Kurdistan Region, Iraq, during two growing season (2020 -2021) to study the effect of Bio health with three concentrations (0, 6 and 12 g.L -1 ), Tecamin max with three concentration (0, 5 and 10 ml.L -1 ), Benzyl adenine with three concentration (0, 100 and 200 mg.L -1 ) and their interaction on vegetative growth characteristics of sweet orange seedling, which were brought from private nursery at Duhok city and had two years old and nearly uniform in growth vigor.All seedlings were transferred from plastic bags to the pots with a capacity of 10 kg mixture of 1:3 organic manure and river sand.The decided concentration of Bio health was dissolved in a liter of water, and then the solution was added to the soil around the seedlings in 20 March for both seasons and was repeated one month later.The previous decided conc. of the Tecamin max and Benzyl adenine were prepared then the seedlings were sprayed as a foliar application three times per season: first, 24/3 /2020 for Tecamin max and 27 ∕ 3 / 2020 for Benzyl adenine and was repeats three time in interval one month for the two growing seasons, the spraying was done till the runoff, Tween-20 was used to increase the surface tension of leaves, the control plants were sprayed only by distilled water with Tween-20.The experiment was arranged in a randomized complete block design with four replications and three seedlings for each experiment unit.All the results were analyzed statistically by using SAS programs (29).Duncan 's multiple range test (DMRT) at 5% level of portability was used to compare the treatments means according to the (5).During the study period, attention was paid to the operations of servicing seedlings, such as irrigation, control of diseases and insects, and removal of weeds.The lath house was also covered with plastic during the cold winter months, from December until March, to protecting the seedlings from frost.RESULTS AND DISCUSSION Plant hight: Data in Table 1 Clearly shows that soil application of Bio health at both concentrations significantly increase seedling high compared to the control, the high value (18.91 and 10.78 cm) for both seasons respectively were obtained in seedling treated with 12 g.L -1 , whereas the less increase in seedling high (13.09and 6.85 cm) were obtained in untreated seedling.Data in same table indicate that spraying of amino acid (Tecamin max) at both concentrations during both seasons led to increase seedling hight.The highest value (18.72 and 10.55 cm) for both seasons respectively were show in seedling sprayed with 10 ml.L-1 of Tecamin max compared to the lowest values showed in control.For the effect of Benzyl adenine, Table (1) appears that spraying sweet orange seedling significantly increased seedling hight in both seasons, maximum increase (18.90 and 10.61 cm) for both seasons respectively were resulted in seedling sprayed with 200 mg.L -1 Benzyl adenine compared to minimum increase (13.45and 7.09) for both seasons respectively were obtained in control.Regarding the interaction among the a above factors same table clearly shows that the best increase (24.250

Stem diameter
Data in Table (2) shows that soil application of Bio health and spraying of Tecamin max and Benzyl adenine all at highest concentration (12g.L -1 ,10 ml.L -1 and 200 mg.L - 1 ) respectively led to increase in stem diameter of sweet orange seedling, the highest increases in stem diameter (2.00, 1.75,1.76,1.70 and 1.75,1.44mm) for both season respectively were produced from application of highest concentration of all studies factors compared to the lowest increase (1.20, 0.79,1.46,0.84 and 1.48, 1.12 mm) for both seasons respectively were obtained in un treated seedling.Concerning the combination effect of studies factor, the best increase in stem diameter were resulted from the interaction among the high concentrations of both Bio health, Tecamin max and Benzyl adenine compared to the less result were obtained in untreated seedling.

Branch numbers
Data in Table (3) shows that the branches number per seedling was significantly stimulatory by soil application of Bio health in different concentrations in both seasons, when compared with untreated seedlings, the highest values (15.34 and 15.22) for both seasons respectively provided when orange seedlings treated with 12 g.L -1 of Bio health, whereas the lowest value of branches number from (12.89 and 6.87) was recording untreated seedlings in both growing seasons respectively.The effect of Tecamin max its notice in same Table that seedling gave the highest value (15.34 and 12.79) when seedlings were treated with 10 ml.L -1 in the two seasons.Same Table shows that the Sweet orange seedlings treated with 200 mg.L -1 Benzyl adenine in both seasons produced the significant highest number of branch when compared with other treatments, which were (16.04 and 12.13) respectively in both seasons and the lowest value (11.90 and 9.13) recorded in untreated seedling in both seasons.Concerning the interaction among the three studded factors, the maximum number of branches (18.250 and 18.375) for both seasons respectively was resulted from the interaction among 12 g.L -1 Bio-health, 10 ml.L -1 Tecamin max and 200 mg.L -1  Benzyl adenine compared to minimum number of branches (8.167&4.833)for both season respectively was resulted in untreated seedling.Tecamin max and 200 mg.L -1 Benzyl adenine respectively for both seasons, as for the combination effect among application of the three factors, data noticed that the maximum leaf area (44.136 and 47.632 cm 2 ) was obtained from the interaction of 12g.L -1 Bio health+5ml.L -1 Tecamin max + 100 mg.L -1 Benzyl adenine in first season and from the interaction of 6g.L -1 + 10ml.L-1 Tecamin max + 200 mg.L -1 Benzyl adenine .and the minimum leaf area (25.925 and 24.553 ) recorded in untreated seedlings Total chlorophyll (µg.m 2-1 ) Data in Table (5) illustrates that the positive effect of all treatments on the chlorophyll content in sweet orange seedling whether added to the soil as Bio health or foliar application as Tecamin max and Benzyl adenine in both seasons, where the highest value of chlorophyll (208.32 and 208.04) that was produced from application Bio-health to the soil at 12g.L -1 in both seasons.Also, the seedling had the highest value of seedling chlorophyll content when treated with Tecamin max and Benzyl adenine at 10ml.L -1 and 200 mg.L-1 respectively, where the values included it (207.11 and 200.6, 206.4 and 198.12) respectively in both seasons.Interaction effect among the Bio health or foliar application as Tecamin max and Benzyl adenine, highest chlorophyll content (215.384 and 216.058) was obtained from the interaction of high concentration of these factors in first season and from the interaction of 12g.L -1 + 0 ml.L -1 Tecamin max + 0 mg.L - 1 Benzyl adenine in second season.

Leaves carbohydrate
According to the data in Table (6) shows that the application Bio health to the soil and foliar application of seedling by Tecamin max and Benzyl adenine especially with in high concentrations led to increase the leaves carbohydrate of sweet orange seedlings when compared with control, the best increase of carbohydrate found in seedlings when treated with 12g.L -1 Bio health as it (9.68 and 7.36) respectively in both seasons, and the seedlings treated with 200 mg.L -1 benzyl adenine gave the highest value (10.32 and 6.625) respectively for both seasons compared with control which produced less values in both seasons..  8) that, at the soil application of Bio health at both concentrations significantly increase leaves dry weight when compared with control, the highest value (0.783, 0.837) g for both seasons respectively were obtained in seedlings treated with 6g.L -1 in first season and 12g.L-1 in second season, whereas the lowest value in leaves dry weight (0.593, 0.719) g were recorded in untreated seedlings.Concerning to the effect of Tecamin max of leaves dry weight of sweet orange seedlings in same Table found that the significant effect of Tecamin max when compared with control, the highest value (0.754, 0.831) g respectively in both seasons recorded when treated with 10 ml.L -1 Tecamin max.As for the effect of Benzyl adenine on the leaves dry weight it was found that non-significant effect was recoded for both concentrations when compared with the control.Leaves dry weight was found to be influenced by the interactions among the studies factors in same Table show the best increasing leaves dry weight (0.886) g was found in seedling that treated with (6g.L -1 , 10ml.L -1 and 0mg.L -1 ) that included Bio-health, Tecamin max and Benzyl adenine respectively in first season, in the second season (1.072) g was recorded in seedlings treated with (12g.L -1 , 10ml.L -1 and 0mg.L -1 ) respectively included bio health, Tecamin max and Benzyl adenine .The lowest value (0.496, 0.582) g respectively in both seasons that in untreated Sweet orange seedlings.The improvement of all the vegetative growth characteristics of local Sweet orange seedlings as a result of soil application of Bio-health , especially at a concentration of 12 g.L -1 , could be attributed to its content of humic acid, seaweed extract and microorganisms, as humic acids contribute to stimulating growth through the content of organic compounds, amino acids, and mineral elements, especially potassium , which actively participates in many physiological processes, such as regulating the work of stomata, as its accumulation in the guard cells affects the osmotic pressure.The process directly affects the water relations inside the plant, including the absorption of water and nutrients from the soil (10,22,23).Humic acid contributes to activating the formation of chlorophyll pigment and the assembly of sugars, amino acids and enzymes (14), activating cell division, increasing the growth rate, developing the vegetative and root system, and increasing the dry matter in plant tissues (35).As for the role of seaweed extract in improving the vegetative growth of seedlings, it could be attributed to the inclusion of many nutrients such as vitamins and amino acids in its composition, which have a wide range of impact on the vital activities of the plant (2,26) as well as "containing plant hormones, especially Auxins and cytokinin's that have an active role in increasing growth and stimulating plant height and side branching (34), and cytokinin's play a major role in stimulating the biosynthesis of chlorophyll in mature tissues, regulating the distribution of nutrients, opening stomata, and delaying leaf senescence (13).The positive role which Tecamin max has played is perhaps due to what it contains of amino acids which reached up to (23) amino-acids, that were necessary for plant growth, and which stimulate metabolism processes which in turn stimulate growth due to the production of necessary growth proteins, vitamins, and enzymes (11,15), or perchance the increase is due to amino-acids working on supplying plants with nitro substances needed for growth and vital processes, which include the division and elongation of plant cells (18).The superiority of the high concentration of Benzyl adenine in the characteristics of vegetative growth could be attributed to the effect of this compound in increasing the strength of the leaves, branches and main stems, which increases the strength and speed pull of processed food and nutrients (4,29), so Its growth increases, in addition to its role in regulating various processes such as cell division and differentiation, stimulating cell expansion and the movement of nutrients to a large degree (16), which affects the growth and development of the seedling, also contributes to controlling the effectiveness of the apical and lateral meristems, which leads to the emergence of new branches as well as delaying the aging of leaves through protecting the photosynthetic pigments from breakdown as well as the differentiation of chlorophyll (20).