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Effect of Surfactants on Bioherbicidal Activity of Alternaria helianthi on Multiple-Seeded Cocklebur



D. Sanyal, P.C. Bhowmik and H.K. Abbas
 
ABSTRACT

Multiple-seeded cocklebur (Xanthium strumarium L.) is a biotype which has different morphology and higher seedling production ability than common cocklebur. Greenhouse studies were conducted to investigate the bioherbicidal activity of Alternaria helianthi (Hansf.) Tubaki and Nishih. on multiple-seeded cocklebur as affected by various rates of Tenkoz COC® (crop oil concentrate), Activator 90® (non-ionic surfactant), BAS 9050 0 S® (methylated oil), Silwet L-77® (organosilicone surfactant) and Top film® (natural based surfactant). Taking X as the recommended rate for each surfactant, 0-X, ¼-X, ½-X, X and 2-X rates were used for each of the surfactants. Surfactants were added to the conidial suspension of A. helianthi. Each surfactant rate was also applied with sterile water without any A. helianthi spore. Treated plants were kept in the dew chamber for 6 h before transferring to the greenhouse. At the end of the experiment (two weeks after treatment), plant shoots were clipped at the soil surface and the fresh weights were determined. Alternaria helianthi resulted in significant reduction in fresh weight of multiple-seeded cocklebur as compared to the plants treated without the fungus. Among five surfactants, Activator 90® and Silwet L-77® had significant effects on the bioherbicidal activity of A. helianthi in reducing fresh weight of multiple-seeded cocklebur. Fresh weight decreased with increase in surfactant rates. Our data demonstrate that A. helianthi control multiple-seeded cocklebur more with higher rates of Activator 90® and Silwet L-77®.

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  How to cite this article:

D. Sanyal, P.C. Bhowmik and H.K. Abbas, 2008. Effect of Surfactants on Bioherbicidal Activity of Alternaria helianthi on Multiple-Seeded Cocklebur. Plant Pathology Journal, 7: 104-108.

DOI: 10.3923/ppj.2008.104.108

URL: https://scialert.net/abstract/?doi=ppj.2008.104.108

INTRODUCTION

Cocklebur (Xanthium strumarium L.) is an economically important weed of soybean [Glycine max (L.) Merr.], cotton (Gossypium hirsutum L.) and peanut (Arachis hypogaea L.) (Byrd and Coble, 1991; Royal et al., 1997; Rushing and Oliver, 1998). Bloomberg et al. (1982) showed that heavy infestation of cocklebur resulted in yield reduction of 50 to 75% in soybean. In addition to the substantial yield reduction, this weed is becoming a concern because several biotypes are resistant to some conventional herbicides. Abbas et al. (1996) showed 12 different biotypes of cocklebur, in which the biotypes from Bolivar county, Mississippi, are resistant to imazaquin (2-[4,5-dihydro-4-methyl-4-(1-methylethyl)-5-oxo-1H-imidazol-2-yl]-3-quinolinecarboxylic acid) and the biotypes from Duck Hill county, Mississippi, are resistant to MSMA (monosodium methylarsonate). A common cocklebur biotype resistant to imidazolinone herbicides was found by Barrentine (1994). Haighler et al. (1988) identified a biotype resistant to the organic arsenical herbicides.

Multiple-seeded cocklebur is a particular biotype of cocklebur, which has a different leaf and stem morphology as compared to the common cocklebur. The burs of the multiple-seeded cocklebur are large, round, covered with hairy spines or prickles, with each seed terminated by a beak (Abbas et al., 1999). This biotype has up to 25 seeds per bur, usually producing up to nine seedlings, whereas common cocklebur has two seeds per bur and usually produces only one seedling (Abbas et al., 1999). Higher seedling production ability increases the weediness of this particular biotype (Abbas et al., 1999; Barrentine, 1974; Bloomberg et al., 1982; Buchanan and Burns, 1971; Snipes et al., 1982).

As some biotypes of cocklebur have become resistant to the conventional herbicides (Barrentine, 1994; Haighler et al., 1988, 1994), the need for an alternative to the chemical weed control methods like biological control has become obvious. Alternaria helianthi (Hansf.) Tubaki and Nishih. has been well documented as a potential bio-control agent for cockleburs (Abbas and Egley, 1996; Abbas and Barrentine, 1995; Quimby, 1989). This is a pathogen of sunflower (Helianthus annus L.) and can infect other plants in compositae family (Allen et al., 1983).

The role of surfactants in increasing the activity of plant pathogens in biological weed control has been well documented (Boyette et al., 1996; Connick et al., 1990; Daigle and Connick, 1990; Watson and Wymore, 1990). Surfactants may improve leaf wettability, improve spore deposition and retention and prolong water retention to overcome dew period requirements (Green et al., 1998). However, there are no guidelines for selecting the compatible surfactants for bio-control agents. Thus, there is a need to investigate the compatibility of surfactants for different pathogen-weed systems.

In the present study, the objective was to determine the effect of five surfactants at various rates on A. helianthi activity on multiple-seeded cocklebur.

MATERIALS AND METHODS

General: Seeds of multiple-seeded cocklebur were collected from plants growing at experimental plots at the Southern Weed Science Research Nursery, Stoneville, Mississippi, established from the seeds from Bell County, Temple, Texas, Collected in 1995. The burs were soaked in water for a week before planting in a 1:1 potting mix of jiffy mix and soil, (Jiffy Mix, Jiffy Products of America, Inc., Batavia, IL 60510). Germinated seeds were transplanted to 10 cm2 pots and grown in the greenhouse (28 to 33° C, 40 to 78% relative humidity and 12 h day length) until 6 to 8 leaf stage.

Preparation of fungal culture: Fungal cultures were taken from a stock culture of A. helianthi and inoculated in fresh sunflower leaf-agar medium prepared by the procedure described by Abbas and Barrentine (1995). Dried sunflower leaves (25 g) were homogenized with 1 L of distilled water in a blender at high speed for 5 min. The homogenate was centrifuged for 10 min at 10,000 x g and the supernatant was filtered through a double-layer cheesecloth. The filtrate was combined with 20 g of agar, autoclaved and poured into 9 cm petriplates, 20 mL each. The plates were inoculated with A. helianthi and incubated at 18°C in alternating regimes of 14 h of fluorescent light at 165 E m-2 sec-1 and 10 h of dark period. After 10 to 14 days of incubation, 5 mL of autoclaved distilled water was added to each plate and the conidia of A. helianthi were scraped from each plate and collected in autoclaved distilled water. Conidial suspensions were homogenized by a polytron PT3000 (Brinkmann Instruments, Inc., Westbury, NY) and counted in a haemocytometer. The concentration of the conidial suspensions were adjusted to 1x105 conidia per mL.

Effect of surfactants on bioherbicidal activity: Effect of five surfactants on the bioherbicidal activity of A. helianthi were studied on multiple-seeded cocklebur. The surfactants used in the present study were Tenkoz COC® (crop oil concentrate), Activator 90® (non-ionic surfactant), BAS 9050 O S® (methylated oil), Silwet L-77® (organosilicone surfactant) and Top film® (natural based). Taking X as the recommended rate for each surfactant, we used 0-X, ¼-X, ½-X, X and 2-X rates for each of the surfactants. The actual rates used were 0, 0.25, 0.5, 1 and 2% (v/v) for Tenkoz COC®; 0, 0.06, 0.125, 0.25 and 0.5% (v/v) for Activator 90®; 0, 0.25, 0.5, 1 and 2% (v/v) for BAS 9050 O S®; 0, 0.05, 0.1, 0.2 and 0.4% (v/v) for Silwet L-77®; and 0, 0.06, 0.125, 0.25 and 0.5% (v/v) for Top film®. Each surfactant rate was applied without and with A. helianthi. Surfactants were added to the conidial suspension before spraying. The six to eight-leaf seedlings were sprayed using a Spra-Tool (Crown Industrial Products Co., Hebron, IL, USA). The treated plants were subjected to 6 h of dew periods before transferring to the greenhouse. Plants were watered and fertilized (with N:P:K, 20:20:20) as needed. At the end of the experiment (two weeks after treatment), plant shoots were clipped at the soil surface and the fresh weights were determined. Experimental designs were completely randomized for all experiments and each treatment had three replications. Data were subjected to analysis of variance (ANOVA) using the general linear model procedures of the Statistical Analysis System (SAS, 1999).

RESULTS AND DISCUSSION

In all experiments, A. helianthi resulted in severe damage and fresh weight reduction of multiple-seeded cocklebur plants as compared to the treatments without the fungus and the fresh weight reduction was significant at 99% level of significance as indicated by the P values in the ANOVA (Table 1). The disease symptoms appeared on infected plant leaves and stems as soft necrotic lesions within 24 h of treatment. Necrotic lesions became dry and larger with time, resulting in death of some plants within 1 week depending on severity of symptoms. These data confirm the effectiveness of A. helianthi as a bioherbicidal pathogen for multiple-seeded cockleburs. This agrees with the findings of Abbas et al. (2004) where A. helianthi caused severe disease infestation and growth reduction of multiple-seeded cockleburs.

Table 1: ANOVA on the effect of various surfactants and Alternaria helianthi on fresh weight of multiple-seeded cocklebur
aA = A. helianthi (with and without); S = Surfactant rates

p-values in the ANOVA show that among five surfactants, Activator 90® and Silwet L-77® had significant effects on the bioherbicidal activity of A. helianthi in reducing fresh weight of multiple-seeded cocklebur (Table 1). Tenkoz COC®, BAS 9050® and Top Film® did not have any effect on fresh weight. Alternaria helianthi reduced the fresh weight of cocklebur plants more at higher rates (0.5%) of Activator 90® as compared to treatments with lower rates (0.06%) or without surfactant (Fig. 1). With Silwet L-77®, A. helianthi resulted in lower fresh weight of multiple-seeded cocklebur plants with higher surfactant rates as well (Fig. 2). Fresh weights were significantly lower at 0.2 and 0.4% rate of Silwet L-77®, as compared to 0.05% rates. At 0.4% rate of Silwet L-77® phytotoxicity was observed on control plants (plants treated with 0.4% Silwet L-77 in sterilized water without any fungal conidia) within 24 h of treatment (Fig. 3). Walker and Tilley (1997) observed similar phytotoxicity due to 0.4% rate of Silwet L-77® on sicklepopd. Previous research also showed that A. helianthi when applied with Silwet L-77® may cause up to 100% mortality of normal and multiple-seeded cockleburs (Abbas et al., 2004).

Overall, A. helianthi resulted in better control of multiple-seeded cockleburs in terms of fresh weight when applied with Activator 90® or Silwet L-77® as compared to Tenkoz COC®, BAS 9050® and Top Film®. Our data demonstrate that with 6 h dew period, A. helianthi reduced the fresh weight of multiple-seeded cocklebur more with higher rates of Activator 90® and Silwet L-77®. Enhanced bioherbicidal activity by specific adjuvants was previously reported by Babu et al. (2003). They showed that the application of Alternaria alternata in an oil emulsion enhanced disease incidence on waterhyacinth (Eichhornia crassipes) as compared to a 0.1% solution of Tween 80®.

Activator 90® and Silwet L-77® have shown potential for achieving effective biological control of multiple-seeded cockleburs by A. helianthi. The precise mode by which these surfactants enhance the activity of A. helianthi is yet to be determined. Surfactants may modify leaf wettability causing fungal conidia to adhere more closely to leaf tissues than fungal conidia in water. Specific surfactants may prolong water retention to overcome dew period requirements as suggested by Green et al. (1998).

Fig. 1: Effect of various rates of Activator 90® (% v/v) on fresh weight of multiple-seeded cocklebur treated with and without Alternaria helianthi two weeks after treatments. Vertical bars represent standard error of means

Fig. 2: Effect of various rates of Silwet L-77® (% v/v) on fresh weight of multiple-seeded cocklebur treated with and without Alternaria helianthi two weeks after treatments. Vertical bars represent standard error of means

Silwet L-77® may stimulate spore germination of A. helianthi, allowing conidia to produce multiple germ tubes to penetrate stomata or possibly wounds caused by Silwet L-77® as well. Abbas and Egley (1996) showed that germination and germ tube production of A. helianthi on cocklebur leaves increased with addition of Silwet L-77® or corn oil. Previous research demonstrated that Silwet L-77® promoted the activity of fungi and bacteria on the leaves of their host kudzu (Pueraria lobata) (Boyette et al., 2002; Zidak et al., 1992).

Multiple-seeded cocklebur has characteristics that suggest that it might be difficult to control, particularly the fact that it produces numerous seedlings per bur, giving the possibility of rapid population growth. However, the results of the current study demonstrate that multiple-seeded cocklebur is highly susceptible to A. helianthi and its effectiveness can be enhanced by use of appropriate surfactants in proper rates with a minimum dew period. As these experiments were tested in the greenhouse, more research is needed in field situation for better understanding of the feasibility of this approach.

Fig. 3: Effect of Silwet L-77® at 0.4% (left) and 0.2% (right) rates (v/v) on multiple-seeded cocklebur 24 h after treatment

Research to enhance the activity of A. helianthi against other biotypes of cocklebur is also needed.

ACKNOWLEDGMENTS

We thank B.J. Johnson for valuable technical assistance. This research was partially supported by the Massachusetts Agricultural Experiment Station, Manuscript No. 3413.

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