Bacteria associated with golden pompano (<Emphasis Type="Italic">Trachinotus blochii</Emphasis>) broodstock from commercial hatchery in Malaysia with emphasis on their antibiotic and heavy metal resistances

This paper was the first report on pathogenic bacteria isolated from golden pompano (Trachinotus blochii) with the antibiogram, as well as the heavy metal resistance pattern. Golden pompano becomes popular among Malaysian fish farmers due to its high value and demand from local and oversea markets. However, the baseline information on antibiogram of pathogenic bacteria associated with golden pompano is not well established. Therefore, the information from this study may be useful to fish farmers in selecting appropriate antibiotic for treatment and prophylactive purpose in golden pompano culture. Isolation of bacterial isolates was carried out using 5% of Horse Blood agar, Tryptic Soy Agar (TSA), Mac Conkey, Thiosulphate Citrate Bile Salt (TCBS), Eosin Methylene Blue (EMB), Glutamate Starch Pseudomonas (GSP), Xylose Lysine Deoxycholate (XLD), and Baird Parker media. The bacterial isolates were then identified using conventional biochemical tests and confirmed by commercial bacterial identification kit. Antibiotic susceptibility test of bacterial isolates against 21 antibiotics (oxolinic acid 2 μg, ampicillin 10 μg, erythromycin 15 μg, furazolidone 15 μg, lincomycin 15 μg, oleandomycin 15 μg, amoxicillin 25 μg, colistin sulphate 25 μg, sulpha-methoxazole 25 μg, chloramphenicol 30 μg, doxycycline 30 μg, florfenicol 30 μg, flumequine 30 μg, kanamycin 30 μg, nalidixic acid 30 μg, novobiocin 30 μg, oxytetracycline 30 μg, tetracycline 30 μg, nitrofurantoin 50 μg, fosfomycin 50 μg, and spiramycin 100 μg) was determined using disk diffusion method, whereas the heavy metal resistance pattern (mercury Hg²⁺, cadmium Cd²⁺, chromium Cr^{6 +}, and copper Cu²⁺) of the bacterial isolates was characterized using two-fold agar dilution method. Five bacterial species were successfully found from 50 diseased golden pompano. They were Streptococcus spp. (n = 12), Escherichia coli (n = 30), Salmonella spp. (n = 20), Pseudomonas spp. (n = 36), and Vibrio spp. (n = 50). More than 80% of the bacterial isolates were found sensitive to 11 out of 21 antibiotics (tetracycline, nitrofurantoin, florfenicol, chloramphenicol, oxytetracycline, nalidixic acid, doxycycline, furazolidone, flumequine, fosfomycin, and oxolinic acid. However, all bacterial isolates were found resistant to all tested heavy metals except for copper and cadmium. The multiple antibiotic resistance (MAR) index of the present study indicated that the fish samples were under high risk exposure to the tested antibiotics. Florfenicol is suggested to be used as antimicrobial agent for golden pompano culture since all bacterial isolates were sensitive to it.     

Photosynthetic ability of Horse gram [<Emphasis Type="Italic">Macrotyloma uniflorum</Emphasis> (Lam.) Verdc] plants in response to metal stress

The effects of heavy metals' stress like aluminium (Al), bismuth (Bi), lithium (Li) and nickel (Ni) on photosynthetic ability of Horse gram plants inoculated with the four metal tolerant rhizobial strains were studied. Horse gram plants can grow in Al contaminated soils up to 400 μg g^–1, at this concentration also they have shown maximum PS ability than in uninoculated plants when they were inoculated with the strain HGR-13. In soils containing Bi, the rate of PS increased up to 200 μg g^–1 with the strain HGR-13 and 400 μg g^–1 with the strain HGR-25. Li has shown significant changes on PS ability of horse gram plants. The PS ability of these plants increased with the strain HGR-6 up to 30 μg g^–1 only. The plants inoculated with the strains HGR-4, 6 and 25 along with uninoculated plants have shown their maximum PS ability at 80 μg g^–1 of Ni. The net PS ability of HGR-4 and HGR-6 was maximum than uninoculated plants. In conclusion, these plants were able to grow in Al, Bi, Li and Ni contaminated soils. The metal tolerant rhizobial strains enhanced the PS ability of horse gram plants when they were inoculated in metal contaminated soils. Horse gram plants and the rhizobia associated with it besides having the nitrogen fixing ability, also have heavy metal tolerance. So, it is concluded that horse gram plants can be used as reclamation agents and phytoremediation.     

Screening and identification of antagonistic <Emphasis Type="Italic">Streptomyces</Emphasis> spp. against <Emphasis Type="Italic">Clavibacter michiganensis</Emphasis> subsp. <Emphasis Type="Italic">michiganensis</Emphasis> from tomato rhizosphere

The objectives of our present study were to isolate antagonistic Streptomyces from tomato rhizosphere, and evaluate the potential strain for the biological control of bacterial canker of tomato. One hundred and seventy strains of isolated from tomato rhizosphere were tested for antibiosis activity against Clavibacter michiganensis subsp. michiganensis on double-layer agar. Sixty-three isolates showed antibiosis activity with diameter of an inhibition zone ranging from 1.0–6.5 cm. Fifteen Streptomyces strains had strong antibiosis activity against C. m. subsp. michiganensis with diameter of the inhibition zone above 4.0 cm on double-layer agar. Especially, the strain named Z-L-22 showed the strongest antibiosis activity with 6.5 cm inhibition zone. The fermentation filtrate also showed a high inhibition activity against Gram-positive bacteria such as Streptomyces scabies, Staphylococcus aureus, and Bacillus subtilis. Morphological, physiological and biochemical tests combined with 16S rDNA sequence analysis were carried out to identify the strain Z-L-22. Characteristics of the Z-L-22 were similar to those of Streptomyces setonii, and the 16S rDNA sequence showed 99.4% homology to S. setonii. Based on the polyphase taxonomic views, the Z-L-22 was identified as S. setonii.     

Presence of indigenous endophytic bacteria in jujube seedlings germinated from seeds <Emphasis Type="Italic">in vitro</Emphasis>

The presence of indigenous endophytic bacteria in tissue-cultured seedlings germinated from seeds of Zizyphus jujuba var. Fupingdazao was investigated using cultivation, light microscopy and scanning electronic microscopy (SEM). In addition, bacteria specific 16S rDNA PCR followed by denaturing gradient gel electrophoresis (DGGE) was used. No cultivable bacteria could be detected on plates or in liquid cultures. However, large quantities of endophytic bacteria in jujube seedlings were observed under the light microscope. The bacterial cells were round (measuring 1.5 μm–1.8 μm), short rods (2.2 μm − 2.9 μm × 1.1 μm − 1.9 μm) and rod shaped (3.7 μm − 4.4 μm × 1.6 μm − 1.9 μm). Rod-shaped bacterial cells (measuring 3.5 m−4.0 m × 1.5m−2.0 m) were observed by SEM as well. A 16S rDNA fragment of 1.5 kb could be amplified from the total DNA of seedlings of jujube using the bacterial primer pair 27F/1525R. The V3 fragment of 16S rDNA was separated by DGGE, and the 16S rDNA-PCR-DGGE showed that there were at least six dominant bands within the seedlings of Z. jujuba var. Fupingdazao. It can be concluded that endophytic bacteria that cannot be detected by cultivation are present with high densities in jujube seeds.     

Growth and yield response of wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) to phosphobacterial inoculation

In this study growth and yield response of wheat crop to phosphobacterium inoculum was observed under sandy loam conditions. The investigations were carried out at field experiment. The experiment was laid out in Randomized Complete Block Design. The treatments were; 120-0-0 NPK kg/ha⁻¹ (T ₁), 120-50-0 kg/ha⁻¹ (T ₂), 120-100-0 NPK kg/ha⁻¹ (T ₃), T ₁ + Phosphobacterium inoculum (T ₄), T ₂ + Phosphobacterium inoculum (T ₅) and T ₃ + Phosphobacterium inoculum (T ₆).The results showed that bacterial strain (Pseudomonas spp.) was able to effect on yield and its attributes in wheat crop. The crop showed significant positive results. The inoculation significantly stimulates the germination count (m⁻²), number of tillers and spikes (m⁻²), 1000 grains weight (g) and grain yield (kg/ha⁻¹). We suggest that application of 120-100-0 kg/ha⁻¹ NPK along with coating of seed with phosphobacterium (Pseudomonas spp.) all the way through inoculation is a better practice to reduce the exploit of phosphatic fertilizers which are much costly.     

Impact of calcium chloride concentrations and storage duration on quality attributes of peach (<Emphasis Type="Italic">Prunus persica</Emphasis>)

To investigate the impact of CaCl₂ concentrations and storage duration on quality of peach (Prunus persica), a research was conducted at postharvest Laboratory, Department of Horticulture, The University of Agriculture, Peshawar, Pakistan during 2012–2013. The experiment was laid out in Completely Randomized Design (CRD) with factorial arrangement repeated three times. The peach fruits (cv. Texas A 69) were harvested at physiological maturity stage from peach orchard, Horticulture Farm. The fruits were dipped in 0, 2 and 4% CaCl₂ solution for 10 min and transferred to cold storage having ±8–10°C with relative humidity of 80–85%. The application of CaCl₂ solution and storage duration significantly influenced the fruit quality of peach fruit. However, the application of CaCl₂ solution significantly reduced weight loss (4.98%), disease incidence (2.08%), total sugars (5.31%), TSS-Acid ratio (16.27), TSS(7.38⁰Brix) and increased the fruit firmness (2.21 kg cm^–2) titratable acidity (0.47%) and Ascorbic acid (5.35 mg/100 g) of peach fruits. The storage duration of peach fruit also significantly affected the fruit quality attributes during storage. The peach fruit stored for 30 days showed less fruit firmness (0.74 kg cm^–2) and titratable acidity (0.31%), Ascorbic acid (4.45 mg/100 g) and increased weight loss (19.74%), disease incidence (16.11%), total sugars (6.07%), TSSAcid ratio (27.62) and TSS(8.54⁰Brix) of peach fruit. Based on the findings of this study, it is concluded that the peach fruit should be treated with 4% CaCl₂ solution to retain the quality attributes for 30 days storage.     

Identification of disease resistances in wheat-<Emphasis Type="Italic">Leymus multicaulis</Emphasis> derivatives and characterization of <Emphasis Type="Italic">L. multicaulis</Emphasis> chromatin using microsatellite DNA markers

To identify resistance to Fusarium head blight (FHB), cereal yellow dwarf virus (CYDV), stem rust (Sr), and powdery mildew (Pm) in 24 common wheat (Triticum aestivum)-Leymus multicaulis addition/translocation lines that were developed cytogenetically and to verify the authenticity of these lines using microsatellite (SSR) DNA markers. Resistance to FHB was identified in the wheat-L. multicaulis addition lines, Line 9 and Line 26, which both contained L. multicaulis-specific fragments as shown by SSR markers. The translocation line, Trans 1, and the addition lines, Line 5 and Line 29, have resistance to stem rust (IT 0). Resistance to CYDV was evaluated based on virus titers measured by enzyme linked immunosorbent assay (ELISA). The addition line, Line 23, showed low virus titer (0.15), indicating resistance to CYDV. The segregation distribution of CYDV resistance in 98 F₂ plants of Line 23/CS showed a significant deviation from 3:1. Inoculation with a set of 14 differential Blumeria graminis f. sp. tritici (Bgt) isolates did not detect powdery mildew resistance in translocation line Trans 1, addition line Line 9 and the amphiploid of wheat-L. multicaulis. However, Line 26 exhibited the resistance response pattern of Kavkaz, which carries Pm8, indicating that Line 26 most likely has the powdery mildew resistance gene Pm8 inherited from its parent lines Feng Kang 7 or Feng Kang 10. Twelve SSR markers, distributed on different homeologous chromosome groups of wheat, which distinguished L. multicaulis addition/translocation chromosomes, were used to verify the presence of L. multicaulis chromatin in the putative wheat-L. multicaulis addition/translocation lines. Of the 24 addition/translocation lines investigated using the 12 polymorphic SSR markers, 18 wheat-L. multicaulis derivatives showed the expected L. multicaulis-specific fragments, indicating that all of these 18 addition/translocation lines would most likely have the introgressed L. multicaulis chromosome(s). Chromosomal rearrangements also were detected in some of the wheat-L. multicaulis introgression lines.     

Sensing soil and foliar phosphorus fluorescence in <Emphasis Type="Italic">Zea mays</Emphasis> in response to large phosphorus additions

Additions of large loads of phosphorus (P) enriched animal manure to soils and the persistence of their environmental impact have been associated with continued water quality impairment in regions of high density of confined animal feeding operations. Foliar P in corn (Zea mays L.) and changes in labile P in Aquic Hapludults were determined following P application of 0–560 kg P ha^?1 as KH₂PO₄ and an application of Fe³⁺ (150 mg Fe³⁺ kg^?1) in field mini-lysimeters to develop calibrations of soil and plant nutritional responses. X-ray fluorescence (XRF) scanning of uppermost leaves of plants at the V2, V5, and V8 stages showed that foliar P proportionally increased with addition rates. Exchangeable and enzyme-labile P forms were effective indicators of foliar XRFS-P for up to 30 days after emergence. Phosphorus calibration curves developed for flag leaves showed that spatial distribution of foliar P (3.6, 4.2, and 5.3 g kg^?1) corresponded to field zones treated with 0, 15, and 30 kg P ha^?1 as dairy manure P for the past 18 years. Up-to-date crop uptake and availability of P in these Hapludults were best described by a square root function of soil XRFS-P and total exchangeable inorganic P (r² = 0.4; RMSE = 419 and 422 g ha^?1, respectively). Therefore, a timely knowledge of canopy P status and its linkage to actual soil P status supports in situ element-specific sensing and precision nutrient management in order to manage the declining use-efficiency in crops and reduce potential loss to the environment.     

Expression analysis of <Emphasis Type="Italic">RUS1</Emphasis> and construction of <Emphasis Type="Italic">RUS1</Emphasis> plant expressing vector

RUS1 was one of the disease resistance gene analogs obtained from Setaria italica Beauv. Semi-quantitative RT-PCR analysis result showed that RUS1 gene could be induced by Uromyces setariae-italicae and had relation to the resistance response of Setaria italica Beauv. against Uromyces setariae-italicae infection. Promoter sequence of RUS1 was obtained by the method of Genome Walking, and its length was 675 bp. RUS1 promoter and pCAMBIA1300 vector were fused to construct RUS1::GUS vector. GUS histochemical staining result showed that promoter could activate gene expression. RUS1 gene (including the promoter sequence) was obtained through PCR amplification and then fused with pCAMBIA1300 vector to construct pCAMBIA1300:RUS1 plant expressing vector. The research laid a foundation for gene functional identification of RUS1.     

Effect of various dietary vitamin A levels on growth performance and immune response of tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>)

Six purified vitamin-free casein-based diets were formulated to contain six levels vitamin A at 0, 500, 1000, 2000, 4000, and 8000 IU·kg⁻¹, respectively. Tilapias (initial mean weight: 7.73±0.03 g) were fed the diets in quadruplicate aquaria to apparent satiation twice daily for 10 weeks. No differences in mortality, weight gain, or feed efficiency ratio (FER) were observed among the groups. Liver vitamin A levels reflected dietary vitamin A levels. Immune parameters, such as hemoglobin levels, total cell count, red blood cell count, total serum protein, and serum lysozyme activity, did not vary with the dietary vitamin A levels. White blood cell counts of fish in 2000 IU·kg⁻¹ diet groups were significantly higher than that in other diets groups. Serum complement activities of fish in 2000 and 4000 IU·kg⁻¹ vitamin A diet groups were also higher than those in other diet groups. After the 14-d challenge test, the mortality and antibody titer were similar among the treatments. The results indicated that dietary vitamin A inclusions did not affect the immune response of Oreochromis niloticus.     

Detection of indigenous endophytic bacteria in <Emphasis Type="Italic">Eucalyptus urophylla in vitro</Emphasis> conditions

The presence of indigenous endophytic bacteria in aseptically grown seedlings of Eucalyptus urophylla germinated from surface sterilized seeds was investigated using dilution plating, microscopy, and PCR detection. No culturable endophytic bacteria could be detected in suspensions of ground plant tissue incubated on solid or in liquid cultivation media. However, a large number of endophytic bacterial cells, mostly rod-shaped and measured 2–3 μm×0.5–0.8 μm, were observed in in vitro cultured seedlings of E. urophylla using both light and electron microscopy. Using the universal bacterial 16S rDNA primers, a predicted 190-bp fragment was amplified from total DNA isolated from the seedlings of E. urophylla. We concluded that the endophytic bacteria originated from the seed were present in seedlings of E. urophylla. However, the bacterial cells observed appeared to be nonculturable.     

Allelopathic effects of <Emphasis Type="Italic">Thymus kotschyanus</Emphasis> on seed germination and initial growth of <Emphasis Type="Italic">Bromus tomentellus</Emphasis> and <Emphasis Type="Italic">Trifolium repens</Emphasis>

The allelopathic influence of aqueous extracts of Thymus kotschyanus on Bromus tomentellus and Trifolium repens germination (%), germination speed, and seedling growth (length, fresh and dry weight) was examined. It was noted that aqueous extracts had a considerable inhibitory effect on target plant germination, and the effect at 50%, 75%, and 100% concentration was found to be significantly higher than that at lower concentrations (5% and 25%) and control treatment (distilled water). Seedling length in addition to fresh and dry weights was also reduced significantly over control. The inhibitory effect was increased as the extract concentration was increased. B. tomentellus showed a higher sensitivity against T. kotschyanus in allelopathic effects compared to T. repens, which indicates that B. tomentellus planted in rangelands with leaf litter of T. kotschyanus will be adversely affected in terms of its germination, growth, and ultimately low forage production.     

Use of molecular markers of potato golden nematode resistance genes <Emphasis Type="Italic">H1</Emphasis> and <Emphasis Type="Italic">GRO1</Emphasis>

By means of DNA markers of potato golden nematode resistance genes H1 and Gro1, 109 Russian- and foreign-bred potato varieties are assessed. A sufficiently high level of coincidence of the presence of marker alleles with phenotypic resistance of potato varieties is revealed. However, not all resistant forms have specific amplification products. The use of DNA markers of only two genes H1 and Gro1 will not make it possible to completely replace mass laboratory testing, but it will make it possible to select potato forms resistant to this parasite by a simple and reliable method in a shorter time and thereby to considerably reduce the number of genotypes in the sample for further breeding.     

Ameliorating Nickel Stress by Jasmonic Acid Treatment in <Emphasis Type="Italic">Zea mays</Emphasis> L.

Plants need various micro and macronutrients for their growth and metabolism. Of these nickel (Ni) plays a role of micronutrient but causes adverse effects when present above optimum level. To combat such situations plants possess different growth regulators. Jasmonates are a class of plant growth regulators modulating various growth and physiological responses in plants. Aiming to evaluate the effect of nickel and jasmonic acid on growth and antioxidant enzymes Superoxide distmutase (SOD), Catalse (CAT), Ascorbate peroxidase (APX), Guaiacol peroxidase (GPX) and glutathione reductase (GR), seeds of Zea mays L. were sown in field after giving presowing treatment of NiSO₄ · 6H₂O (8 mM), jasmonic acid JA (10^–6, 10^–8 and 10^–10 M) alone and JA (10^–6, 10^–8 and 10^–10 M) in 1: 1 combination with NiSO₄ · 6H₂O (8 mM) in plant conservatory, Punjabi University, Patiala, Punjab, India. Plants of each treatment group were then subjected to spraying treatment with the respective treatment solutions at regular intervals of 22 days up to 66 days after sowing (DAS). The present findings signified reduced growth with Ni treatment but co-application of JA alleviated the effect of nickel through activity of antioxidant enzymes.     

Effects of salicylic acid on the behavior of Yali pear infected by <Emphasis Type="Italic">Alternaria kikuchiana</Emphasis> Tanaka

To investigate the inductive effect of salicylic acid (SA) on the resistance of Pyrus bretschneider cv Yali to black spot disease (Alternaria kikuchiana Tanaka), the physiological and biochemical characteristics of detached pear leaves at the age of 5 to 10 days were measured after application of SA. The results showed that exogenous SA significantly improved the resistance of Yali pear (Pyrus bretschneider cv Yali) leaves to black spot disease. For the SA treatment at 0.02 mmol·L⁻¹ SA concentration, the disease index was the lowest, and the induced resistance reached up to 63.9%. Furthermore, SA induced local and systemic resistance of Yali pear against the black spot disease. Expression of systemic resistance in leaves was detectable 3 d after SA treatments and lasted for 10 d. POD, PPO, and PAL activities of Yali pear leaves increased by SA treatment. It is suggested that exogenous SA solution as a chemical activator could induce the resistance of Yali pear to black spot disease.     

Formation of the hypersensitivity response due to the expression of <Emphasis Type="Italic">FeSOD1</Emphasis> gene in tomato when it is inoculated with <Emphasis Type="Italic">Phytophthora infestans</Emphasis>

The activity of antioxidant enzymes and ultrastructural changes in tissues inoculated with P. infestans isolate have been studied in the previously developed independent transgenic lines of tomato with FeSOD1 gene and control plants. It is shown that the activity of superoxide dismutase is significantly higher in transgenic plants than that in control plants (nontransgenic plants). Chlorosis and obvious changes in tissue turgor were observed when the control tomato plants were inoculated, which indicates irreversible damages and unimpeded progression of infection. At the same time, the transgenic lines were characterized by the formation of clearly limited zones of damaged cells that rapidly arrested the infection. In addition, the damages differed from those in nontransgenic plants: the cells along the edges of the infection site were smaller and had heavy invaginations of the cell wall. The contacts between the cells were disrupted in this zone, but they were preserved in undisturbed zones of the tissue. Thus, the expression of the FeSOD1 transgene promotes the emergence of the resistance to P. infestans in tomato transgenic plants.     

MPN-PCR enumeration of <Emphasis Type="Italic">Campylobacter</Emphasis> spp. in raw chicken meats and by-products

The goal of this study is to enumerate Campylobacter in chicken meats and by-products. In the current investigation, results showed that raw chicken meats and chicken by-products were contaminated with Campylobacter ranging from <3 to 4600 MPN·g⁻¹. Campylobacter jejuni showed a higher number compared to Campylobacter coli in the chicken samples. The current study showed that the percentage of chicken livers and gizzards harbored a higher number of Campylobacter (10³–10⁴ MPN·g⁻¹) than other chicken parts at 33.3% and 9.2%, respectively. The different concentrations of Campylobacter between chicken meats and chicken byproducts reflect the differences in the contamination level. The data on Campylobacter concentration in chicken meats and by-products will be useful in risk analysis.     

Characteristics of interspecific hybrids between <Emphasis Type="Italic">Pleurotus ostreatus</Emphasis> and <Emphasis Type="Italic">P. eryngii</Emphasis>

In this study, three species of oyster mushroom (Pleurotus osturatus, P. eryngii and P. cornucopia) were crossed together in order to aggregate benefit special attributes to the genotype (s). The monokaryon of each of these species was prepared. Then, the monokaryons of two species were placed at 5 mm distance from each other to produce dikaryon. The results showed that, from among 700 crosses, only the crosses of eryngii and osturatus monokaryons were grown toward each other and produced clamps (dikaryons). Four produced hybrids were noted H₁, H₃₂, H₁₁ and H₄₀. The spans of produced hybrids were prepared; then, they were grown into sterile media and attributions of hybrids were studied. The results indicated that H₁₁ hybrid was an appropriate hybrid in terms of the number of days until growing, number of days until the observation of the first pin and the days from planting to harvest. However, H₄₀ was the best hybrid based on cap diameter, dry and fresh weight of fruit body, yield and biomass. It is expected that these interspecific hybrids employ for future oyster mushroom breeding programs.     

New strain of <Emphasis Type="Italic">Vicia unijuga</Emphasis> mosaic virus revealed in the Amur oblast

A mosaic disease caused by a virus with isometric particles with a diameter of 28–30 nm has been revealed on a wild vetch species Vicia unijuga in the Amur oblast. The thermal inactivation point (TIP) of the pathogen is 73°C, longevity in vitro (LIV) is up to 3 days, and dilution endpoint (DEP) is 10^?5. Virions contain one capsid protein with molecular mass 19 kD. The antigenic relationship between the isolate and brome mosaic virus and earlier identified V. unijuga mosaic virus, a representative of the genus Bromovirus, is determined. The pathogen has been identified as a new strain of this bromovirus. The virus is easily transmitted to cultivated legumes, causing considerable damage.     

The optimizing conditions of soft-wood cutting of yam-lobation <Emphasis Type="Italic">Atractylodes macrocephala</Emphasis> Koidz.

This study was focused on optimizing the conditions of soft-wood cutting for Atractylodes macrocephala Koidz., including the concentration and treatment time of IBA and IBA + NAA and the proportion of turf to perlite in substrates. L₉(3⁴) orthogonal design with three factors and three levels were adopted, and the best combination was chosen after formula analysis, variance analysis, and range analysis. The results showed that both the maximum survival rate and rooting rate were 93.33%. Statistic analysis showed that treatment with IBA (200 mg·L⁻¹) + NAA (100 mg·L⁻¹) for 20 s, turf to perlite ratio of 1:3 would be the most favorable combination for soft-wood cutting of A. macrocephala.