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Herbicidal Potential of Eucalyptus Dundasii on Lolium Rigidum Gaud. and Hordeum Spp

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Herbicidal Potential of Eucalyptus Dundasii on Lolium Rigidum Gaud. and Hordeum Spp Seventeenth Australasian Weeds Conference Herbicidal potential of Eucalyptus dundasii on Lolium rigidum Gaud. and Hordeum spp. Hanwen Wu1, Jinbiao Zhang1,3, Rex Stanton2, Min An2, De Li Liu1 and Deirdre Lemerle2 1 EH Graham Centre for Agricultural Innovation (Industry and Investment NSW and Charles Sturt University), Wagga Wagga Agricultural Institute, PMB, Wagga Wagga, NSW 2650, Australia. 2 EH Graham Centre for Agricultural Innovation (Industry and Investment NSW and Charles Sturt University), Charles Sturt University, Locked Bag 588, Wagga Wagga, NSW 2678, Australia 3 Test Centre, Fujian Agricultural and Forestry University, Fuzhou, Fujian 350002, PR China Corresponding author: [email protected] Summary Field observation has found that there and Leptospermum myrsinoides. Eucalyptus contains is limited understory vegetation within the dripline a rich source of bioactive constituents, possessing of Eucalyptus dundasii. Research was conducted to fungicidal, insecticidal and herbicidal activities (Zhang assess the allelopathic potential of E. dundasii on et al. 2010). The bioactive compounds derived from two important weeds species, annual ryegrass and eucalyptus have been reported to cause phytotoxicity barley grass. The eucalyptus leaves were collected to a range of field crops, such as wheat (Triticum aes- and subjected to a distillation process. The distillation tivum), maize (Zea mays), radish (Raphanus sativus) process yielded three bioactive fractions, essential oil and rice (Oryza sativa) (Batish et al. 2004, Batish et fraction, water fraction A and water fraction B. These al. 2006, Khan et al. 2008). three fractions were phytotoxic to the germination The phytotoxic effects of eucalyptus allelochem- and seedling growth of the annual ryegrass and barley cials have also been evaluated on a number of weed grass. Essential oils caused the most inhibitory effects species. Aqueous leaf leachate of Eucalyptus globulus on germination and shoot growth of both weeds, fol- suppressed the establishment of vegetative propagule lowed by water fraction B and fraction A. Both weeds and early seedling growth of purple nutsdge (Cyperus had similar responses when exposed to the essential rotundus L.) and Bermuda grass (Cynodon dactylon oils. However, the germination and seedling growth L.) (Chandra Babu and Kandasamy 1997). The her- of barley grass were more sensitive than annual bicidal activity of eucalyptus oils against Parthenium ryegrass to water fraction A and the germination of hysterophorus, Cassia occidentalis, Echinochloa crus- annual ryegrass was more sensitive than barley grass galli and A. viridis has also been documented (Batish et when exposed to water fraction B. The phytotoxicity al. 2004, Batish et al. 2006, Singh et al. 2005). These was concentration-dependent. The research has found results suggest that bioactive compounds in eucalyp- that although essential oils are highly volatile, these tus oils have potential commercial value for further compounds can partly dissolve in water, resulting in exploitation as natural herbicides (Zhang et al. 2010). the phytotoxic activity identified in the water fraction Field observation has found that there is limited A. These results indicate that any significant rainfall understory vegetation within the dripline of Dundas events will wash off the essential oils into the ground blackbutt (Eucalyptus dundasii). This research was to perform natural chemical defense against understory conducted to assess the allelopathic potential of the vegetation, including weeds. Further study on the eucalyptus on two important weeds, annual ryegrass identification of bioactive compounds might provide (Lolium rigidum Gaud.) and barley grass (Hordeum chemical leads for the development of new herbicides spp.). with new modes of action. Keywords Eucalyptus dundasii, allelopathy, MATERIALS AND METHODS weeds, annual ryegrass, barley grass. Plant material Fresh leaves of E. dundasii were col- lected from fields in Ungarie, NSW, Australia. Seeds INTRODUCTION of annual ryegrass were purchased commercially and Eucalyptus is a member of Myrtaceae family and a barley grass seeds were collected locally from Wagga native to Australia. It has long been recognised that Wagga Agriculture Institute research station in 2008. some eucalyptus species are capable of suppressing un- derstorey vegetation growth via allelopathy (May and Steam distillation Fresh leaves (300 g) of E. Ash 1990). Del Moral et al. (1978) demonstrated that dundasii were subjected to steam-distillation for 2.5 Eucalyptus baxteri was capable of allelopathically sup- h using a Pyrex oil distillation apparatus with a flat pressing the understorey growth of Casuarina pusilla bottom flask (2 L) containing 1200 mL distilled water 139 Seventeenth Australasian Weeds Conference to generate steam. The distillation process produced proposed by An et al. (2005). The whole-range as- three fractions: essential oils, water fraction A and sessment considers overall effect/response across the water fraction B. The volatile components from whole range of application rates, instead of assessing leaves were condensed through cooling tubes. The the effect of each individual rate on test species. The essential oil afloat on top of the condensed water program WESIA (Whole-range Evaluation of the was collected through a separation funnel. The cor- Strength of Inhibition in Allelopathic-bioassay) was responding condensed water was also collected and used to calculate the inhibition index based on the designated as water fraction A (full strength, 100%). following equation (Liu et al. 2007): The collected essential oils were stored in a sealed vial at 5°C before use. After steam distillation, the residual water remain- ing in the flat bottom flask turned brown as a result of the reflux of vapour through the eucalyptus leaves contained in the upper distillation flask. This residual water was filtered, collected and designated as water where the 0, D1, D2, … Dn, are the dose-concentrations fraction B (full strength, 100%). The collected water tested and the R(0), R(D1), R(D2), … R(Dn) are the fractions A and B were stored in a freezer prior to use. corresponding responses, respectively. The Dc is the threshold dose at which response equals the value of Bioassays of essential oils on weed germination and control and above which the responses are inhibitory. growth A previous bioassay protocol (Azirak and f(D) represents the response function. Karaman 2008) was adopted with slight modifications. Fifty seeds of annual ryegrass or twenty-five seeds RESULTS of barley grass were separately sown onto 9 cm Petri Germination Results revealed that germination of dishes lined with one layer of Whatman No.1 filter both annual ryegrass and barley grasses were inhibited paper. Distilled water (5 mL) was initially delivered by the essential oils (Figure 1a) and by the two water to each Petri dish. An aliquot of 0, 12.5, 25, 50 or 100 fractions A and B (Figure 1b). The germination of L of E. dundasii essential oil was then added directly the two weeds had similar responses to E. dundasii onto the filter paper to test the inhibitory effect of the essential oils. The phytotoxicity of essential oils was essential oil. Immediately after the treatment, each enhanced by increased amount of essential oils ap- Petri dish with its cover was sealed with a piece of plied. The germination of both weeds was inhibited parafilm to reduce evaporation. All Petri dishes were by more than 90% at the essential oil quantity of 50 maintained in a growth incubator with a diurnal cycle L dishí1. of 30°C with light and 15°C with dark and a photope- The germination of the two weeds responded riod of 12 h. A randomised complete block design differentially when exposed to water fractions A or B. with three replicates was used. Germinated seeds with Water fraction B was more inhibitory than fraction A >1 mm radicle were recorded and shoot lengths meas- (Figure 1b). For water fraction A, barley grass germi- ured after 10 days of incubation. nation was more sensitive than ryegrass germination. However, ryegrass seed germination was more sensi- Bioassays of water fraction A and B on weed germi- tive to water fraction B than barley grass. nation and growth The bioassay protocol developed The analysis of whole-range assessment showed by Wu et al. (2003) was adopted. A concentration that phytotoxicity of the three fractions ranked in de- series was made up from the full strength (100%) creasing order as essential oils, water fraction B and solutions of water fraction A or B into 100%, 75%, water fraction A (Table 1). 50%, 25% and 0% (water control). Fifty seeds of an- nual ryegrass or twenty-five seeds of barley grass were Seedling growth Annual ryegrass and barley grass sown onto 9 cm Petri dishes lined with one layer of had similar responses when exposed to essential oils Whatman No.1 filter paper. An aliquot (5 mL) of each or the water fraction B (Figure 2), while the weeds concentration of water fraction A or B was delivered to responded differentially to the water fraction A. Water each Petri dish. The management of Petri dishes and fraction A was more inhibitory to the seedling growth measurements were as previously described. of barley grass than annual ryegrass. Essential oils caused the most inhibitory effects Data analysis The dose-response data were sub- on shoot growth of both weeds, followed by water jected to the analysis of whole-range assessment fraction B and fraction A (Table 1). 140 Seventeenth Australasian Weeds Conference a. 120 a. 100 100 80 80 60 60 40 40 Growth (%) inhibition 20 ARG Germination inhibition (%) inhibition Germination ARG BAR 20 BAR 0 020406080100120 0 020406080100120 Essential oil concentrations (μL) Essential oil concentrations (μl) b. 100 120 b. 80 100 60 80 40 60 Fr A - ARG Fr A - BAR Germination inhibition (%) inhibition Germination 20 Growth (%) inhibition 40 Fr B - ARG Fr B - BAR Fr A - ARG Fr A - BAR 0 20 020406080100120 Fr B - ARG Fr B - BAR Concentration of water fractions A and B (%) 0 020406080100120 Figure 1.
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