1. Introduction
Cotton (
Gossypium hirsutum L.) is an important commercial crop worldwide, and serves as a significant source of fiber, feed, foodstuff, oil and biofuel [
1,
2]. Cotton is the pillar industry of Xinjiang Uygur Autonomous Region, China, which is of great significance to the economic and social development of Xinjiang [
3]. Defoliation or leaf abscission is induced in cotton as a natural physiological process, which is usually inadequate or not timely enough for a complete mechanical harvest of cotton. Therefore, defoliation before harvest is often induced by managing the plants so that senescence, abscission (separation) layer development and leaf drop are encouraged [
4,
5,
6,
7]. Advantages associated with defoliant application prior to cotton harvest include: increased harvester efficiency, reduction in the leaves and trash content in harvested lint, and quicker drying of dew. Defoliant spraying is a key link in the mechanized cotton harvest, as sufficient and uniform spraying could improve the defoliation quality and decrease the cotton trash content, and it is significant to solve defects of cotton quality [
8,
9]. Mechanization, modernization and standardized production are the basic paths of sustainable development of the Xinjiang cotton industry [
10]. Most cotton pesticide operations are carried out using large-volume ground machinery in Xinjiang, which results in rolling the cotton plant, hitting the bolls, pulling the cotton branch, hitting the opened balls off, water and pesticide wastage, and reduces the yield and quality of cotton. It is a bottleneck and technical issue that restricts the cotton quality and efficiency of Xinjiang. Herbicidal defoliant diuron and hormonal defoliant thidiazuron are widely used in Xinjiang. Thidiazuron increases the concentration of ethylene relative to auxin in leaf petioles and results in the activation of the leaf abscission layer [
11,
12,
13]. Diuron accelerates the scorch of cotton leaves, improves defoliation under low temperature conditions [
14]. However, these types of defoliants induce drastic leaf abscission which inhibits timely transport of nutrients from leaves to cotton bolls. Also, these defoliants do not directly influence boll ripening and must be applied in combination with ethephon, a boll opener, to provide satisfactory defoliation and boll opening [
15,
16].
Plant protection unmanned aerial vehicles (UAVs) are light and small UAVs with pesticide spraying equipment. UAVs achieve precision pesticide spraying using the global positioning system (GPS) and geographic information system (GIS) [
17,
18]. The advantage of UAVs is using low-volume spray technology replaced the traditional large-volume mass locomotive spray technology. [
19]. UAVs accomplish spray precision docking by GPS and real-time kinematic (RTK) automatic navigation technology, which increases the quality of aviation plant protection operations significantly [
20]. Compared to conventional agricultural aircraft, UAVs do not require a special airport and have additional advantages, such as good mobility [
21,
22]. UAVs are also more adaptable for spraying at low altitudes due to geographical restrictions [
23,
24]. Meanwhile, UAVs could effectively improve pesticide efficacy, reduce pesticide amounts, and rational use of pesticide [
25,
26]. The problem of the development of plant protection UAVs is the combination and relationship between aircraft and agricultural plant protection in China [
27,
28]. Plant protection is accompanied by the reduction in agricultural disease, insects, weed control and prevention, while low-altitude aviation is emerging as a breakthrough in aviation technology. The initial combination of aviation and plant protection has resulted in low-altitude aviation plant protection. UAVs have a very important value in the development of precision agriculture, fulfilling an urgent need in the development of modern agriculture in China. Currently, leaves of the cotton canopy overlap, making it inconvenient for crop spraying using a conventional land spraying machine. Due to the harsh walking conditions in cotton fields, operating a land-spraying machine is very difficult and requires high labor intensity. Large-volume spraying not only leads to pesticide waste but also seriously endangers the environment and the operators [
29]. The development of aviation plant protection technology would drive the development of new pesticide industries and the aviation application equipment industries in line with the needs of national strategic development which has broad market prospects and bring about huge economic, social and ecological benefits [
30].
Boom sprayers are widely used for large farm crops due to their high working efficiency and favorable spraying effect. However, there are still some problems in cotton defoliant spraying in Xinjiang. Cotton is planted in a high density in Xinjiang, the row space is 10 cm and 66 cm, causing leaves in two adjacent rows to be seriously overlapped, which results in reducing the yield and quality of cotton [
9]. Zhao et al. [
31,
32] and An [
33] reported control of cotton aphids, Qin et al. reported control of plant hoppers by UAVs, which showed good results and could effectively decrease insect population trends [
34]. However, there are few reports on the spraying defoliants by UAVs. Herein, the authors report this study’s results concerning the effect of defoliant dosage on defoliation, boll opening, absorption and decontamination in cotton leaves and the effect of spraying volume on absorption and decontamination in cotton leaves sprayed by plant protection UAVs, to obtain the optimized parameters for the working of UAVs, and provide references and bases for further improving the cotton defoliant spraying technique.
2. Materials and Methods
2.1. Experimental Material
2.1.1. The UAV Sprayers and Equipment
The aviation platforms were the JT-30 UAV (UAV 1,
Figure 1 and
Figure 2, Xinjiang Jiangtian Aviation Science and Technology Co., Ltd., Xinjing, China) and the 3WQF120-12 UAV (UAV 2,
Figure 1, Anyang Quanfeng Aviation Plant Protection Technology Co., Ltd., Anyang, China), which were equipped with a spraying unit. The main parameters of the UAV 1 and UAV 2 are presented in
Table 1. The UAVs both used GPS and RTK navigation technology, with the accuracy of the flying height and flying velocity controlled within 0.1 m and 0.1 m/s, respectively. The spraying platforms consisted of a medical kit with a capacity of 30 L (UAV 1) and 12 L (UAV 2), a miniature straightway pump, pipeline, spraying nozzle, and electronic control valve. UAV 1 with six spraying nozzles (hollow conical nozzle) were symmetrically arranged on both sides of the UAV at the same interval (700 mm), and the installing angle of the 4 spraying nozzles was a vertically downward spraying direction and using 2 spraying nozzles with a 60° angle in a horizontal direction. UAV 2 with three spraying nozzles (LU120-02) were symmetrically arranged on both sides of the UAV and in the precise middle at the same interval (625 mm), and the installing angle of the spraying nozzles was a vertically downward spraying direction. Using a working pressure of 0.3 MPa (UAV 1) and 0.3 MPa (UAV 2), the measured flow rate of a single spraying nozzle was 250 mL/min (UAV 1) and 400 mL/min (UAV 2).
Thidiazuron and diuron were analyzed with an Agilent 1200 HPLC equipped with a reversed-phase column (Agilent XDB-C18 4.6 × 150 mm, 5 μm, Agilent, Santa Clara, CA, USA) at 25 °C. The mobile phase was methanol/water (65/35, v/v) at a flow rate of 0.8 mL/min. The injection volume was 20 μL and electronic balance, BSA224S-CW, was from Sartorius, Göttingen, Germany. High speed centrifuge, LDZS-2, was from Beijing Jingli centrifuge Co. Ltd., Beijing, China. Swirl meter, MS3 D S25, IKA, came from Staufen im Breisgau, Germany. Ultrasonic cleaner, KQ-500 DB, was from Kunshan Ultrasonic Instruments Co. Ltd., Kunshan, Jiangsu, China and the Rotary Evaporators, RV10 D, were from IKA, Staufen im Breisgau, Germany.
2.1.2. Defoliant and Reagents
80% thidiazuron wettable powder was produced by Jiangsu Repont Pesticide Factory Co., Ltd., Changzhou, China. 37% thidiazuron + 17% diuron suspension concentrate, came from Jiangsu Institute of Ecomones Co., Ltd., Nanjing, China. 40% ethephon aqueous solution produced by Jiangsu Anpon Electrochemical Co., Ltd., Huaian, China were used. Thidiazuron standard (97%) and diuron standard (97%), from J&K Scientific Ltd., Beijing, China were used. Methanol (HPLC), came from Sigma-Aldrich, St. Louis, MO, USA. Primary secondary amine (PSA) sorbent, was produced by Welch Technology (Shanghai) Limited Co., Ltd., Shanghai, China.
2.2. General Situation of Experimental Field
The effect of defoliant dosage on defoliation, boll opening, absorption and decontamination in cotton leaves experiment was carried out in Beiquan town of Xinjiang Production and Construction Crops, Shihezi, (E 85°28′32″, N 44°39′54″), Xinjiang Uygur Autonomous Region, China, during September 2017. The experimental field had middle level fertilizer and planted cotton for many years. Cotton (Xinluzao 64) was planted on 24 April 2017, using a mechanical cotton-picking planting model, with wide film planting 6 lines (10 cm + 66 cm), 195,000 cotton /ha, and drip irrigation under plastic film. The effect of spraying volume on the dynamics of defoliant absorption and decontamination in cotton leaves experiment was carried out in Farm 150 of Xinjiang Production and Construction Crops (E 86°03′28″, N 44°56′42″), Shihezi, Xinjiang Uygur Autonomous Region, China, during September 2017. The experimental field had middle level fertilizer planted cotton for many years. Cotton (Xinluzao 64) was planted on 21 April 2017, using a mechanical cotton-picking planting model, a wide film planting 6 lines (10 cm + 66 cm), 180,000 cotton /ha, drip irrigation under plastic film. The heights, growing ways and leaf area index were uniform within each farm. The difference of the two farms was the density, which was affected by the soil fertility and planting density.
2.3. Experimental Treatment
2.3.1. Effect of Defoliant Dosage on Defoliation Effect Sprayed by UAV 1
The experiment consisted of 4 treatments (
Table 2). Cotton defoliant spraying was carried out on 7 September 2017 and 15 September 2017.
2.3.2. Effect of Spraying Volume on Defoliation Effect Sprayed by UAV 2
The experiment consisted of 4 treatments (
Table 3 and
Table 4). Cotton defoliant spraying was carried out on 5 September 2017 and 13 September 2017. The weather during the experimental period is in
Table S1.
2.4. Defoliation and Boll Opening
Prior to treatment application, 30 plants were randomly tagged to count the number of leaves on each plant. The number of leaves was counted again 4, 8, 12 and 15 day after spraying on the same tagged plants. Defoliation rate was calculated by Equation (1).
where N
a = Number of leaves before treatment, N
b = Number of leaves after treatment.
Boll opening rates were determined on the same tagged 30 plants. Bolls on each plant were examined and recorded as either opened or closed and the boll opening rate was calculated by Equation (2).
where N
c = Number of opened bolls, N
d = Number of Total bolls.
2.5. Cotton Leaves Processing
2.5.1. Cotton Leaves Extraction and Purification
The cotton leaves (upper, middle and lower layer) were collected before spraying and 1, 4, 7, 8 and 12 days (1, 5, 7, 8 and 15 days for spraying volume experiment) after spraying. There are eight fruit branches per cotton, the upper, middle and lower layers are three, three, and two fruit branches, respectively (
Figure 3). A method had been developed to determine defoliant residues in cotton leaves using QuEChERS-pretreatment method and HPLC. Sample preparation and purification were performed as follows: 4.0 g of cotton leaves were accurately weighed, frozen in liquid nitrogen and grinded into powder. The powder was transferred to a 50 mL centrifuge tube. Afterwards, 25 mL of acetonitrile was added to the centrifuge tube, and the mixture was ultrasonically extracted for 20 min. Subsequently, NaCl (3 g) was added to the solution, which was vortexed for 1 min and centrifuged at 3800 rpm for 5 min. Next, 1 mL of supernatant was transferred into a 2 mL centrifuge tube with 50 mg of PSA, vortexed for 1 min and centrifuged at 10,000 rpm for 3 min. The supernatant was filtered through a 0.22 μm organic membrane filter for HPLC. Blank samples were used for validation studies and matrix-matched standard calibration. An untreated group was set up as control group, and all samples were stored in a −20 °C freezer.
2.5.2. Standard Curve and Added Recovery
The thidiazuron and diuron standard (50 mg, respectively) were dissolved in 50 mL methanol and diluted to 100 mL together, obtaining 500 mg/L thidiazuron and diuron mixed mother liquid, then diluted to 2, 4, 6, 8 and 10 mg/L, respectively, resulting in a series of mixed standard solutions. The standard solution was determined by HPLC. The standard curve was obtained by drawing the area as an ordinate and drawing the concentration of the thidiazuron and diuron standard solution as abscissa. The standard curve equation of thidiazuron and diuron were y = 29.845x − 23.63, R2 = 0.9964 and y = 114.33x − 82.01, R2 = 0.995, respectively. The peak area and concentration of thidiazuron and diuron exhibited a good linear relationship (2–10 mg/L).
Samples for recovery studies were spiked with the corresponding volume of the working solution and incubated for 30 min before extraction. Cotton leaves were treated with 0.05, 0.5 and 5 mg/kg (thidiazuron) and 0.05 and 0.5 (diuron) standard working solution, and thidiazuron and diuron recoveries were determined using an established method. Each additional level was repeated three times whilst setting a blank control (
Table 5 and
Table 6). The average recoveries of 0.05, 0.50 and 1.0 mg/kg thidiazuron from cotton leaves were 81.47%, 83.11% and 93.71%, respectively. The average recoveries of 0.05 and 0.50 mg/kg diuron from cotton leaves were 117.28% and 100.75%, respectively. These values met the requirements for residue analysis.
2.6. Yield Characters and Fiber Quality
The cotton yield was measured after all the cotton bolls opened where 100 cotton bolls from the canopies (upper, middle and lower layer) were randomly tagged and collected in each experimental area to determine the cotton yield characteristic and fiber quality.
2.7. Data Statistics and Processing
Defoliation, boll opening, uniformity and elongation of the cotton fiber which were expressed in percentage, were transformed to arcsin√X/100; others were log (x + 1) transformed prior to analysis to stabilize wide variances and meet normal assumptions. After transformation, the data was analyzed for normality using the Kolmogorov-Smirnov test and for equal variance across the treatments and replicates using Levene’s test. Data was compared across different application rate using analysis of variance (ANOVA) (SPSS v21.0, SPSS Inc, an IBM Company, Chicago, IL, USA). Duncan’s new multiple test was used for multiple comparisons and the significance level was p = 0.05. In the factorial design for both experiments, the averages were compared by using t-test at 5% probability.
4. Conclusions
In conclusion, the pooled data from this study indicate that plant protection UAVs could be used for cotton defoliants spraying by a twice defoliants spraying strategy, the first spray composed of 80% thidiazuron, the second spray the compounded formulations 37% thidiazuron + 17% diuron, which could guarantee the defoliation and boll opening effectively. The defoliant dosages had no significant effect on seed cotton yield and fiber quality. Meanwhile, the residue of thidiazuron in cotton leaves reached the maximum four days after spraying and the residue of diuron in cotton leaves reached the maximum one day after the second spraying. The thidiazuron and diuron residues increase with spraying volume in the range of 17.6–29.0 L/ha. When the spraying volume is less than 17.6 L/ha, the residue of thidiazuron and diuron is reduced. According to the experiment results and combined with the cotton defoliation, boll opening, fiber quality and thidiazuron and diuron residues in cotton leaves, the first spraying dosage of 300 g/ha and spraying volume of 17.6 L/ha will be recommended to the farmers for UAVs defoliant application. The research results could provide a reference for further optimization of the spraying parameters of cotton defoliant by plant protection UAVs.