Fenazaquin resistance development and management in two spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae)

doi:10.21203/rs.3.rs-2458609/v1

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Fenazaquin resistance development and management in two spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae)

https://doi.org/10.21203/rs.3.rs-2458609/v1

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Fenazaquin resistant population of two-spotted spider mite, Tetranychus urticae (Koch) was developed by giving selection pressure. After 8 generations, the resultant population of mite obtained having resistance ratio of 216.74 folds (FR-strain) as compared to susceptible population (S-strain) maintained in the laboratory. FR-strain was found to have resistance breakdown upto 158.82-times when selection pressure was withdrawn for three generations. Developmental stages of fenazaquin resistant strain were bigger in size and took lesser number of days to complete incubation period, development of immature stage and total life cycle than in susceptible strain. However, fecundity and adult longevity of S-strain was more than FR-strain. The FR-strain was examined for cross resistance against different acaricidal products, namely, hexythiazox (5.45 EC), propargite (57 EC), azadirachtin (0.15 EC), Darekastra and Tamarlassi. These resulted in lower value of resistance ratio, being < 2 suggesting their incorporation in managing fenazaquin resistant population of T. urticae.

Entomology

Tetranychus urticae

Fenazaquin

resistance development

cross resistance

IRM

Two-spotted spider mite (TSSM), Tetranychus urticae Koch (Acari: Tetranychidae) is one of the most important mite pests in the world affecting wide range of field, vegetable, fruit and ornamental crops (Ghongade and Sood 2018; Chouikhi et al. 2022; Ghongade et al. 2022). It is well adapted to various environmental conditions, causing loss of quality, yield, and death of plants by sucking out the contents of leaf cells. Indirect effects of mite feeding include decreased photosynthesis and transpiration. It has a rapid process of host range evolution as reported by Gould (1979). Tetranychus urticae is considered as one of the “most resistant species” in terms of total number of pesticides to which populations have become resistant (Leeuwan et al. 2010). The species is known to rapidly develop resistance to acaricides due to its polyphagous nature, short life cycle, high fecundity, arrhenotokous parthenogenesis and very broad host range (Adesanya et al. 2021).

Intensive application of acaricides has been used to control the TSSM, which ultimately result in acaricide resistance in field populations globally. Abamectin resistance ranged high to very high and milbemactin resistance level were found low in European field population by Xue et al. (2020). Zhang et al. (2022) found TSSM field population in China to be highly resistant to abamectin, pyridaben, profenofos, bifenthrin. Sun et al. (2022) evaluated laboratory built resistant strain of cyetpyrafen exhibiting resistance ratio of > 2000 fold and found that Cyet-R showed serious cross resistance to cyflumetofen (190 folds) and to cyetpyrafen (> 2500 folds). Salman and Saritas (2014) developed a laboratory resistant strain of TSSM with High resistance level of 122 folds. Abamectin selected resistance population with resistance level of 342- folds was found unstable when no selectin pressure was given by Sato et al. (2005).

The chemical treated females of two spotted spider mite had shorter longevity and lower reproductive period as reported by Kim et al. (2006), Marcic et al. (2012), Li et al. (2017) and Havasi et al. (2018). In the laboratory study described here, we conducted studies on development of Fenazaquin resistance strain of Tetranychus urticae by giving selection pressure, determining resistance breakdown in Fenazaquin resistant strain and its cross resistance to different acaricides. Life cycle and growth of susceptible and resistant strain of Tetranychus urticae were also examined.

Tetranychus urticae population was raised under controlled environment at 25+1 ^oC temperature, 70+5 per cent relative humidity and photoperiod of 16 hrs (16L: 8D) over excised mulberry and French Beans leaves. These leaves were kept over moist sponge sheets in trays.

Test acaricides: Fenazaquin (Magister 10 EC) was used to develop a resistant strain of Tetranychus urticae. Synthetic acaricides namely, Hexythiazox (5.45 EC), Propargite (57 EC), biopesticide, azadirachtin (0.15 EC) and natural products namely, Darekastra and Tamarlassi were used to evaluate the cross-resistance spectrum in the fenazaquin resistant strain

Development of susceptible strain: The mites were raised without exposing them to any chemicals for almost 25 generations and thus, the population obtained was designated as susceptible stain (S-strain).

Development of Fenazaquin resistant strain: Lethal toxicity to susceptible mite strain was determined by using leaf dip method of bioassay as suggested by Erdogan et al. (2012) and is designated as parental generation (F₀). In this, one day old adult female mites (n= 30) were gently released over the treated excised mulberry/ French Bean leaf discs (dia 3cm) with different concentrations of fenazaquin with the help of camel hair brush. After 24 hrs mortality and LC₅₀value were calculated. The per cent mortality was calculated as:

The corrected mortality was calculated as per the method suggested by Abbott (1925).

After obtaining LC₅₀ value, about 500 adult females of F₀ generation were treated with the LC₅₀ value. The surviving individuals were shifted to new untreated leaves after 24 hrs for obtaining next generation and this generation was regarded as F₁ Generation. The adult female mites from the F₁ generation were further bioassayed with same procedure till appreciable increase in resistance to fenazaquin resulting in extremely high resistance was obtained. Then, it was designated as fenazaquin resistant strain (FR-strain) as per the method suggested by Kim et al. (2004).

The data obtained were subjected to probit analysis as per Finney (1971) and LC₅₀ value along with upper and lower fiducial limits were calculated for different acaricidal products. The LC₅₀ values for all the chemical acaricides were calculated and toxicity was considered to be significantly different when 95 per cent confidence limit failed to overlap.

Growth and development of Fenazaquin Resistant Strain (FR-Strain): The observations on size and duration of different developmental stages of FR- and S-strain were recorded. The adult female mites (n=10) of FR- and S-strain were released separately on the leaf discs (dia: 3cm) with the help of camel hair brush and replicated five times. After 24 hrs, the eggs laid by mites on each leaf disc were counted, after the adult mites were removed under stereomicroscope. These were reared to adult stage. When the 50 per cent of individuals in a replication were transformed to larva and adults the time elapsed in between were established as incubation and total developmental period. Measurements on each developmental stage were made for both the strains using imaging software (Nikon NIS elements). Adults were transferred to new leaf discs (five pairs) to record their longevity and fecundity and replicated thrice.

Stability of resistance in FR-strain: The adult females of T. urticae of FR-strain (F₈ generation) and S-strain were bioassayed and multiplied up to three generation without applying fenazaquin selection pressure under laboratory conditions. Similarly, the adult females of S-strain were also bioassayed. The LC₅₀ values for each generation were determined for FR- and S-strain of T. urticae and assessed as per the procedure elaborated earlier. Resistance ratio was worked out as per the formula described earlier and deviation in the level of resistance in FR-strain after non-selectivity for three generations was determined.

Cross resistance spectrum of FR-strain: The fenazaquin resistant strain (FR-strain) developed by giving selection pressure upto eight generations was examined for cross resistance against chemical acaricides (propargite and hexythiazox), biopesticide (azadirachtin) and natural products (Tammarlassi and Darekastra). Leaf dip method of bioassay described previously as suggested by Erdogan et al. (2012) was used. Simultaneously, the adult females of S-strain were bioassayed to compare the variation in toxicity level. Further, the LC₅₀values of different acaricides/ products to FR- and S- strain were worked out. And resistance ratio was worked out as per the formula given hereunder:

Statistical analysis: The data obtained were subjected to statistical analysis as suggested by Gomez and Gomez (1984) using different statistical tools.

Development of Fenazaquin resistant strain of Tetranychus urticae

The mortality in parental generation (F₀) ranged from 16.7 to 83.3 per cent in concentration range of 0.0003 to 0.005 per cent. The obtained LC₅₀ value was 0.0012 per cent with the fiducial limits of 0.0008–0.0016 per cent. When individuals of parental generation were given the selection pressure of 0.0012 per cent of fenazaquin, it resulted in LC₅₀ value of 0.0019 per cent in F₁ generation of FR-line. Whereas, the LC₅₀ value for S-strain was 0.0011 per cent. The difference in toxicity level of fenazaquin in F₁ generation of FR-line and S-strain were non-significant. The F₁ generation exihibited the resistance factor of 1.72. After fenazaquin selectivity of F₁ population, the LC₅₀ value computed for F₂ was 0.0042 per cent. The toxicity value (LC₅₀) obtained for F₂ generation of FR-line was significantly higher to S-strain (0.0013%). In generation F₂, the resistance ratio rose to 3.33. With the increase in selection pressure of fenazaquin concentration to 0.0042 per cent, the LC₅₀ value for FR-strain in F₃ raised to 0.0076 per cent. The LC₅₀ value obtained for S-strain (0.0012%). In F₃ generation of FR-line, the level of resistance became 6.14. The third generation (F₃) of FR-line when exposed to fenazaquin selection pressure (0.0076%) resulted in the LC₅₀ value of 0.0136 per cent. For S-strain, the resultant LC₅₀ value was 0.0011 per cent. In this generation (F₄), the resistance ratio reached 12.04 times with respect to S-strain depicting moderate level of resistance as per method suggested by Kim et al (2006). For obtaining F₅ generation of FR-line, adult females from F₄ generation were treated with fenazaquin concentration (0.0136%). Further, bioassay of the multiplied individuals in F₅ generation resulted in LC₅₀ value of 0.0377 per cent. In contrast to it, LC₅₀ value obtained for S-strain was 0.0013 per cent. The resistance level reached to moderate level of resistance with corresponding value of 30.19. The LC₅₀ value obtained for sixth generation of FR-line was 0.0616 per cent. In contrary to it, the LC₅₀ for S-strain was 0.0011 per cent. The resistance ratio obtained for FR-line and S-strain was 54.00 representing high level of resistance. When selection pressure of 0.0616 per cent given to individuals of F₆ generation of FR-line, the LC₅₀ value obtained for F₇ generation was 0.1475 per cent. For susceptible population, the LC₅₀ value obtained remained almost identical to previous generations (0.0013%) However, the resistance ratio reached to 117.05 being high. With further fenazaquin selection pressure (0.1475%) to adult females of F₇ generation of FR-line, the LC₅₀ value for F₈ generation became 0.2601 per cent, whereas, the LC₅₀ value obtained for S-strain was 0.00120 per cent. In this generation, the resistance ratio reached 216.74 being extremely high.

The LC₅₀ value for FR-line increased from initial level of 0.0012 per cent recorded in parental generation (F₀) surged to 0.2601 per cent over eight generations by inducing fenazaquin selection pressure as represented in Fig. 1 and Table 1. The median toxicity values of FR-line and S-strain were on a par in F₁ generation only, becoming significantly higher in FR-line during F₂ generation onwards. Toxicity level of fenazaquin in F₀ and F₁ generation of FR-line was on par to each other. The toxicity level of fenazaquin increased significantly in subsequent generation being on par in F₂ & F₃, F₄ & F₅, F₅ & F₆ and F₇ & F₈ generations.

Table 1

Toxicity of fenazaquin to adult females of fenazaquin resistant (FR-) line and susceptible (S-) strain of *Tetranychus urticae* in successive generations under selection pressure
Generation	Line/ Strain	Regression equation (y)	LC₅₀(%)*		Resistance ratio
Generation	Line/ Strain	Regression equation (y)	LC₅₀	Fiducial limits	Resistance ratio
Parental (F₀)		3.3784 + 1.5280 x	0.00115	0.00080 - 0.00164
F₁	FRL	2.7129 + 1.7855 x	0.00191	0.00137 - 0.00256	1.72
	SS	3.2780 + 1.6499 x	0.00111	0.00079 - 0.00152
F₂	FRL	2.4858 + 1.5502 x	0.00419	0.00291 - 0.00582	3.33
F₂	SS	3.3154 + 1.5325 x	0.00126	0.00089 - 0.00178
F₃	FRL	2.3199 + 1.4244 x	0.00761	0.00501 - 0.01082	6.14
F₃	SS	3.3862 + 1.4742 x	0.00124	0.00087 - 0.00178
F₄	FRL	2.1049 + 1.3568 x	0.01360	0.00849 - 0.01956	12.04
F₄	SS	3.4481 + 1.4734 x	0.00113	0.00078 - 0.00161
F₅	FRL	2.5685 + 1.5420 x	0.03774	0.02687 - 0.05275	30.19
F₅	SS	3.2342 + 1.6078 x	0.00125	0.00090 - 0.00175
F₆	FRL	3.8299 + 1.4824 x	0.06156	0.04142 - 0.08656	54.00
F₆	SS	3.3278 + 1.5821 x	0.00114	0.00081 - 0.00159
F₇	FRL	3.2233 + 1.5202 x	0.14748	0.10337 - 0.20777	117.05
F₇	SS	3.2560 + 1.5870 x	0.00126	0.00090 - 0.00176
F₈	FRL	3.0934 + 1.3473 x	0.26009	0.16899 - 0.37713	216.74
F₈	SS	3.2424 + 1.6270 x	0.00120	0.00087 - 0.00166

Level of resistance was recorded low in F₁, F₂ & F₃ generations (≤ 10), moderate in F₄ (12.04) & F₅ (30.19) and at high level for F₆ (54.00) & F₇ (117.05). The resistance ratio reached extremely high for F₈ generation corresponding to 216.74. Resistance ratio obtained in generations (F₁ to F₈) was used to develop a predictive model for assessing the resistance ratio in subsequent generations. An exponential regression model fitted best to the data with R² value of 0.9982 and resulted in the equation, y = 0.8045e^0.7035x; where, y is resistance ratio, e is exponential constant and x is generation number (Fig. 2). The resistance ratio of 216.74 was achieved by selection pressure in eight generations. However, Sharma et al. (2018) reported the resistance ratio to be 166.49 after fifteen generations of selection pressure. The variation is attributed to the difference in methodology for giving the selection as they gave selection pressure to two subsequent generations after F₂ up to F₁₅.

Resistance strain of TSSM was also developed for other acaricides under laboratory conditions. After 40 applications of clofentezine, extremely high resistance was observed in mite population in Australia (Herron et al. 1993). Kim et al. (2006) observed extremely high resistance ratio (RR = 252) after selection of 20 generations for fenpyroximate in Korea. Resistance ratio of 274 to spirodiclofen under laboratory selection was observed by Pottelberge et al. (2009). Sato et al. (2016) observed the resistance ratio of after 20 selections in TSSM for spiromesifen from Brazil.

Stability of resistance in FR-strain of Tetranychus urticae

Studies on stability of resistance to fenazaquin in T. urticae strain incorporated by selection pressure during eight generations were undertaken by rearing FR-strain without any selection pressure for three generations. When fenazaquin selection pressure was abstained for one generation, the LC₅₀ value obtained was 0.2412 per cent for NSF₁ with resistance ratio of 191.4. For S-strain, the LC₅₀ value of 0.0013 per cent. Without giving any selection pressure of fenazaquin further for two generations, the LC₅₀ of 0.2190 per cent was obtained, whereas, in S-strain the LC₅₀ value obtained was 0.0013 per cent. The resistance level remained extremely high being 173.80. Additional non-selection of NSF₂ for next generation, the LC₅₀ value obtained was 0.1938 per cent. For S-strain, the LC₅₀ value obtained was 0.0012 per cent. The resistance ratio declined to 158.82 and categorized as high in NSF₃.

There is breakdown of resistance in FR-strain and the median lethal concentration (LC₅₀) ranged from 0.1938 to 0.2412 per cent. Whereas, for S-strain, the LC₅₀ value remained stable (0.0012–0.0013%). The resistance ratio of 191.4 and 173.8 was obtained for NSFG₁ and NSFG₂, respectively being extremely high, whereas, it declined to high level (158.8) for NSFG₃. The per cent reduction in resistance owing to non-selectivity during three generations was 11.68, 19.81 and 26.72 per cent, respectively when compared to baseline resistance ratio of F₈ generation (216.74).

Resistance ratio obtained in NSFG₁ to NSFG₃ was used to develop a predictive model for determining the complete breakdown of resistance in FR-strain. A linear regression model fitted best to the data with the R² value of 0.984 and resulted in the equation, y= -19.13x + 233.0; where, y is resistance ratio and x is generation number.

Table 2

Breakdown in resistance to fenazaquin in adult females of fenazaquin resistant (FR-) strain of *Tetranychus urticae* after generation gap of non-selection pressure
Generation	Strain	Regression equation (y)	LC₅₀		Resistance ratio	Per cent breakdown in resistance
Generation	Strain	Regression equation (y)	LC₅₀	Fiducial limits (%)	Resistance ratio	Per cent breakdown in resistance
NSFG₁	FR-	3.2805 + 1.2439 x	0.24119	0.14732 - 0.35890	191.42	11.68
	S-	3.4456 + 1.4130 x	0.00126	0.00087 - 0.00184
NSFG₂	FR-	3.0477 + 1.4565 x	0.21899	0.14270 - 0.30771	173.80	19.81
	S-	3.1543 + 1.6752 x	0.00126	0.00092 - 0.00174
NSFG₃	FR-	2.8979 + 1.6330 x	0.19376	0.12960 - 0.26430	158.82	26.72
	S-	3.1847 + 1.6715 x	0.00122	0.00088 - 0.00167

Baseline resistance ratio value for FR-strain = 216.74

The resistance was found to be unstable in FR-strain. The instability in resistance without any selection pressure was evident from the findings of Flexner et al. (1995) and Nicastro et al. (2009) for organotin and milbemectin resistance in TSSM. Sato et al. (2016) also observed the resistance to spiromesifen being unstable as it decreased from 75 to 15 per cent in span of 6 months in the absence of selection pressure. However, Nicastro et al. (2013) reported the chlorfenapyr resistance being stable in the absence of selection pressure under laboratory conditions in T. urticae.

Growth and development of FR-strain of Tetranychus urticae

Morphometric Parameters

It is observed that developmental stages of FR-strain except the egg stage were significantly bigger in size as compared to S-strain. The larva, protonymph and deutonymph of FR-strain measured 193, 261 and 341µm, respectively, being 9.04, 10.59 and 9.65 per cent bigger than susceptible strain. Even, the adult males and females of FR-strain were 8.23 and 5.86 per cent bigger in size to S-strain and measured 342 and 524µm in length, respectively.

Table 3

Morphometrics of different developmental stages of FR- and S-strains of *Tetranychus urticae*
Developmental stage	Length (mean ± SE, µm)		t_calculated
Developmental stage	FRS	SS	t_calculated
Egg	126.16 ± 3.66	125.28 ± 2.45	0.449
Larva	193.60 ± 15.36	177.18 ± 11.00	1.710*
Protonymph	261.58 ± 13.63	236.93 ± 16.61	2.566*
Deutonymph	341.11 ± 13.54	311.29 ± 16.63	2.829*
Adult male	342.81 ± 16.53	316.22 ± 15.87	3.464*
Adult female	524.27 ± 13.43	495.15 ± 22.47	2.508*
*Significant at t_tab (0.05) (df: 28)

A literature surveyed revealed no such information. However, in present studies the findings of other workers are being compared to S-strain. The size of eggs of S-strain of T. urticae recorded in present studies is in close proximity to that observed by Devi (2011), Kaur (2014) and Patil et al. (2014) who reported it to be 130, 135.4 and 130µm, respectively. The length of larva recorded in present studies (177.1µm) was bigger to those reported by Devi (2011) and Kaur (2014) who reported it to be 160 and 152.5µm, respectively. The size of the T. urticae male is in close proximity to that revealed by Mallik and Channabasavanna (1983) and Kaur (2014), whereas found smaller as per the findings of Devi (2011) and Patil et al. (2014) and females were found bigger in size in present findings.

Developmental biology and fecundity of FR- and S-strain

It is observed that fenazaquin resistant strain took less time to complete different developmental stages of its life cycle as compared to S-strain and resulted in lower fecundity. The duration of egg stage differed significantly and was 2.2 and 2.8 days in FR-strain and S-strain, respectively. Total duration of immature stages (larva, protonymph and deutonymph) was 9.0 and 9.4 days in respective strains, however, the variations being non-significant. The total developmental period from egg to adult stage was 10.8 and 12.2 days for FR-strain and S-strain, respectively, differing significantly at p = 0.05. Observations recorded that females of FR-strain were short lived (8.4 days) as compared to S-strain (9.0 days). The females live longer as compared to males. Whereas, the male longevity was 5.6 and 6.4 days in FR- and S-strain, respectively. Female mites of FR-strain laid significantly less number of eggs 51.40 eggs/ female than the S-strain 56.00 eggs/ female.

The finding for incubation periods as recorded by Kaur (2014) was 2.52 days, Kaur and Zalom (2018) was 3.33 days and Devi (2011) was 3.9 days. The duration taken by immature stage was 4–8 and 7.53 days as per the findings of Devi (2011) and Kaur (2014). Female longevity recorded by Devi (2011) and Kaur (2014) was more 7–11 & 10.5 and 6.7 & 9.4 days, respectively.

Table 4

Development biology and fecundity of FR- and S- strain of *Tetranychus urticae*
Stage	Mean ± SE		t_calculated
Stage	FRS	SS	t_calculated
Egg (duration in days)	2.20 ± 0.55	2.80 ± 1.40	2.385*
Immature stages (duration in days)	9.00 ± 1.75	9.40 ± 2.08	0.214
Egg to adult emergence (duration in days)	10.80 ± 1.03	12.20 ± 1.04	8.210*
Adult longevity (days)
Male	5.60 ± 0.67	6.40 ± 1.42	3.499*
Female	8.40 ± 1.41	9.00 ± 1.24	3.385*
Total fecundity (number of eggs/ female)	51.40 ± 3.98	56.00 ± 2.78	26.616*
*Significant at t_tab (0.05) (df: 8)

However, the fecundity rate of T. urticae reported by Liu (1989), Puttaswamy (1978), Devi (2011), Patil et al. (2014) and Kaur and Zalom (2018) was 71, 45.5, 70–135, 76 and 44.75 eggs, respectively. Incubation period reported by Aswathi and Bhaskar (2014) was 2.92 days and adult longevity was 12 and 12.5 days in male and female, respectively. Pottelberge et al. (2009) found total fecundity (after 24 hrs) in susceptible and spiromesifen-resistant strain of two spotted spider mite being 60 and 62 eggs, respectively.

Pesticidal exposure is reported to reduce the duration of life cycle as well as fecundity as documented by Kim et al. (2006). They reported the exposed females of T. urticae to have lower longevity, reproductive period and fecundity. Marcic et al. (2012) reported in spirotetramat selection pressure net fertility reduces to 12.4–88.8 per cent with reduced female longevity when compared to control. Li et al. (2017) also observed reduced survival rate (9 and 13%), oviposition period (77.6 and 83.1%), fecundity (89.2 and 76.9%) and longevity (79.2 and 83.1%) in TSSM when exposed to bifenazate at LC₁₀ and LC₂₀ level of selectivity.

Cross resistance spectrum of FR-strain

The FR-strain of T. urticae developed giving selection pressure for eight generations resulting in resistance level of 216.74 fold was evaluated for cross resistance to different acaricidal products and the findings are being presented in Table 5.

Azadirachtin

The observation revealed that azadirachtin concentrations ranging from 0.00002 to 0.00045 per cent for FR- and S-strain resulted in LC₅₀ values obtained were 0.00013 and 0.00011 per cent, respectively. The resistance ratio obtained for FR- strain was low (1.18).

Table 5

Cross resistance against different products to fenazaquin resistant (FR-) strain and susceptible (S-) strain
Product name	Strain	Regression line (y = a + bx)	LC₅₀ (%)		Resistance Ratio
Product name	Strain	Regression line (y = a + bx)	LC₅₀	Fiducial limit	Resistance Ratio
Acaricides
Hexythiazox*	FR-	3.3276` + 1.8601 x	0.00079	0.00065–0.00097	1.11
Hexythiazox*	S-	3.6266 + 1.6092 x	0.00071	0.00055–0.00092	1.11
Propargite	FR-	3.6352 + 1.5997 x	0.00713	0.00517–0.01011	1.03
Propargite	S-	3.8749 + 1.3385 x	0.00693	0.00473–0.01051	1.03
Biopesticide and Natural products
Azadirachtin	FR-	3.4070 + 1.4180 x	0.00013	0.00009–0.00020	1.18
Azadirachtin	S-	3.5204 + 1.4263 x	0.00011	0.00007- 0.00016	1.18
Darekastra	FR-	2.8174 + 1.3303 x	4.37206	3.08343–6.41331	1.10
Darekastra	S-	3.3375 + 1.0404 x	3.96264	2.53959–6.40521	1.10
Tamarlassi	FR-	3.0144 + 1.2768 x	3.59102	2.46409–5.25237	1.15
Tamarlassi	S-	3.4514 + 1.0349 x	3.13623	1.94365–4.90043	1.15
*Eggs were treated

Darekastra

The LC₅₀ values obtained for FR-strain and S-strain of T. urticae were 4.37206 and 3.96264 per cent, respectively for the evaluated concentrations of 0.62 to 20.00 per cent. The resistance ratio obtained for Darekastra was 1.10.

Hexythiazox

For hexythiazox, the egg stage was bioassayed in concentrations ranging between 0.00017 to 0.00272 per cent. For FR-strain, the LC₅₀ value obtained was 0.00079 per cent. The median lethal concentration value obtained for S-strain was 0.00071 per cent resulting in the resistance ratio of 1.11.

Propargite

The adult females of T. urticae evaluated against propargite resulted in LC₅₀ values for FR- and S-strain to be 0.00713 and 0.00693 per cent. Low level of cross resistance was observed with corresponding value of 1.03.

Tamarlassi

The concentrations at which the product was evaluated varied from 0.62 to 20.00 per cent. The median lethal concentration for Tamarlassi was 3.59102 and 3.13623 per cent for FR- and S-strain. It resulted in the resistance ratio of 1.15.

The LC₅₀ value for different acaricidal products varied from 0.00013 to 4.37206 per cent for FR- strain and 0.00011 to 3.96264 per cent for S-strain. The cross resistance ratio varied from 1.03 to 1.18, being minimum and maximum for propargite and azadirachtin depicting it to be of low level for all the acaricides.

Resistant pest population also exhibit varying level of resistance to different pesticides. Kim et al. (2004) observed fenpyroximate resistant population to result in high level of cross resistance to propragite (RR = 64) and low level to fenazaquin. Sato et al. (2005) observed abamectin resistant strain to exihibit no cross resistance towards propargite. Whereas, Salman and Saritas (2014) observed moderate level of cross resistance for hexythiazox in acequinocyl resistant strain. No cross-resistance was found between Cyenopyrafen & SYP-9625 and bifenazate in laboratory strain of Tetranyhus urticae by Chen et al. (2019). No cross resistance was observed by Xue et al. (2020) between abamectin and milbemectin selected population of two-spotted spider mite in European population.

Judicious use of acaricides, utilization of new chemicals with different mode of action and inoooperation of natural products and biopesticides can help farmers to get control of the acaricide resistance in Tetranychus urticae.

The authors declare no competing interests.

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Fenazaquin resistance development and management in two spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae)

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