Journal of Asia-Paci c Entomology · Effects of plant-growth-promoting microorganisms and...

Journal of Asia-Pacific Entomology 17 (2014) 587–593

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Journal of Asia-Pacific Entomology

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Effects of plant-growth-promoting microorganisms and fertilizers ongrowth of cabbage and tomato and Spodoptera litura performance

Yuwatida Sripontan a, Ching-Wen Tan a, Mei-Hua Hung b, Chiu-Chung Young b, Shaw-Yhi Hwang a,⁎a Department of Entomology, College of Agriculture and Natural Resources, National Chung Hsing University, 250 Kuo-Kuang Road, Taichung 402, Taiwanb Department of Soil and Environmental Sciences, College of Agriculture and Natural Resources, National Chung Hsing University, 250 Kuo-Kuang Road, Taichung 402, Taiwan

⁎ Corresponding author. Tel.: +886 422840363; fax: +E-mail address: [email protected] (S.-Y. H

http://dx.doi.org/10.1016/j.aspen.2014.05.0071226-8615/© 2014 Korean Society of Applied Entomolo

a b s t r a c t
a r t i c l e i n f o
Article history:Received 9 December 2013Revised 13 May 2014Accepted 15 May 2014Available online 27 May 2014

Keywords:Plant-growth-promoting microorganisms(PGPMs)FertilizerFoliar chemistryInsect performance

Fertilizer and plant-growth-promoting microorganisms (PGPMs) both benefit crop growth; however, little isknown about the interaction effects when they are combined. This study assessed the effect of PGPMs andfertilizer on plant growth, foliar chemistry, and subsequent insect feeding. Cabbage and tomato plants wereinoculated with PGPMs (fungi and bacteria) and various levels of fertilization. Plant growth parameters (freshweight, dry weight, and leaf area) and foliar chemistry (water content, protein content, and polyphenol oxidaseactivity) were then analyzed. In addition, foliage was also fed to the third instar larvae of Spodoptera litura toevaluate foliage quality. The results indicated that plant performance differed significantly among treatments,and the combined fungiMeyerozyma guilliermondii and fertilizer treatment promoted the greatest plant growth.In summary, PGPMs and fertilization canhave their owneffect; their interaction effect, however, need to be clarified.© 2014 Korean Society of Applied Entomology, Taiwan Entomological Society and Malaysian Plant Protection

Society. Elsevier B.V. All rights reserved.

Introduction

Nearly all plant species undergo various levels of herbivory duringtheir life spans and have developed various strategies to oppose theseattacks (Harrison, 2005; Johnson and Agrawal, 2005; Karban andBaldwin, 1997). They protect themselves against herbivory by using aset of morphological and chemical defense strategies (Dicke andHilker, 2003; Harrison, 2005; Johnson and Agrawal, 2005; Karban andBaldwin, 1997). Chemical defense strategies involve secondarymetabolites and proteins that may be present constitutively or inducedby challenges such as herbivore wounding (Bennett and Wallsgrove,1994; Duffey and Stout, 1996; Ryan, 1990; Zhu-Salzman et al., 2008).Although the evolution of such defense traits can be genetically fixed(Adler et al., 1995; Berenbaum et al., 1986; Hwang and Lindroth,1997), the outcome of such traits might also be amended by otherenvironmental factors (Bryant et al., 1983; Herms and Mattson, 1992).

Various environmental features are considered to affect plants’allocation of resources to defensive compounds (Bryant et al., 1983;Herms and Mattson, 1992). Nutrient accessibility has been considereda vital factor that influences plant growth and the distribution of limitedresources (Bryant et al., 1983;Hemming and Lindroth, 1999). Fertilizationhas been considered a fundamental method of improving soil nutrientavailability for plants and may consequently affect the growth, time ofmaturity, plant part size, and phytochemical content of plants (Altieri

886 4 22875024.wang).

gy, Taiwan Entomological Society an

and Nicholls, 2003; Hemming and Lindroth, 1999; Mevi-Schütz et al.,2003; Myers, 1985). The phytochemical changes caused by fertilizationin host plants may successively affect the pest species that use the hostplants (Altieri and Nicholls, 2003).

Microbial cooperation in the rhizosphere is essential for thesustainability of soil fertilizer and plant growth. Previous literaturehas indicated that mutualistic microbials in soil improve nutrientavailability for plants and provide additional benefits, such as moreefficient uptake of minerals and water, disease resistance, protectionfrom heavy metal toxicity, and an improved soil structure (Clark andZeto, 1996; Nichols and Wright, 2004; Pawlowska et al., 2000).Plant-growth-promoting microorganisms (PGPMs) are defined assoil-borne bacteria and fungi with plant promotion or protectionactivities (Bashan and de-Bashan, 2005; Kamath et al., 2008).PGPMs have been reported to affect plants’ yield, physiology, growthand germination rates, and protein, mineral, and chlorophyll content(Bashan and de-Bashan, 2005; Glick, 2004; Lai et al., 2008). Ingeneral, microorganisms promote plant growth in two processes:(1) PGPMs might fix atmospheric nitrogen to facilitate the absorptionof solubilizing phosphorus and iron as well as to elevate the productionof plant hormones, such as auxins, gibberellins, and cytokinins; and(2) PGPMs might also induce plants’ resistance to phytopathogens(bacteria, fungi, and viruses) (Bashan and de-Bashan, 2005; Figueiredoet al., 2011), insect pests (Ramamoorthy et al., 2001; Zehnder et al.,1997), and nematode pests (Ramamoorthy et al., 2001; Sikora, 1988).Therefore, using selected PGPMs to promote host plants’ resistance toinsect pests may be a potential alternative for sustainable agriculturalpractices.

d Malaysian Plant Protection Society. Elsevier B.V. All rights reserved.

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588 Y. Sripontan et al. / Journal of Asia-Pacific Entomology 17 (2014) 587–593

For example, Zehnder et al. (1997) determined that applying plant-growth-promoting rhizobacteria (PGPR) can induce resistance againstcucumber beetle feeding in cucumbers. The cotton inoculated withPseudomonas gladioli can also reduce the relative growth rate anddigestibility of Helicoverpa armigera because of the induced increase infoliar polyphenol and terpenoid content (Qingwen et al., 1998). Inaddition, one study indicated that, when rice (Oryza sativa) wasinoculated with various PGPR strains, the incidence of rice leaf folder(Cnaphalocrocis medinalis) was reduced (Loganathan et al., 2010)because of a greater accumulation of enzyme lipoxygenase andchitinase activity against leaf folder insects. Therefore, numerousbeneficial soil-borne microbes are suggested to cooperate with plantsto combat insect herbivores by promoting plant growth and inducingresistance (Pineda et al., 2010).

Thus, PGPMs may play particular roles in a plant’s induced defenseresponse. However, such induced responses may vary among abiotic-ecological conditions (Stout et al., 1998; van Dam et al., 2001). Amongthese environmental factors, the extent of resource availability wasobserved to exert a profound impact on the level of the induced defenseresponse (Barto et al., 2008; Borowicz et al., 2003) because severaldefensive substances contained elements derived from nutrients(Herms and Mattson, 1992). The effect of the interaction betweennutrient availability and microbial-mediated induction response hasnot previously been addressed and the effect of the interaction-induced responses on subsequent herbivore performance also remainsunknown. Therefore, this study assessed the interaction effects ofresource availability and PGPMs on plant growth performance andinduced response as well as on the performance of Spodoptera litura.

Materials and methods

Plants

Two plant species, cabbage (Brassica oleracea L. var. capitata L.) andtomato (Lycopersican esculentum Mill. Var. cerasiforme (Dunal) Alef.),were used in the experiment. Seeds were sown with soil (Stenderpeat substrate, Known-You Seed Co., Ltd., Taiwan) in 104-well platesin a greenhouse (25±2 °C, 16:8 h light/dark photoperiod) andwatereddaily. After 2 weeks, the seedlings were transplanted into 12.7 cm potsfilled with field soil. Before planting, the chemical properties of the soilwere analyzed by the Soil Survey and Testing Center, National ChungHsing University, Taichung, Taiwan. Table 1 shows the test results.Forty-seven-day-old cabbage and 51-day-old tomato plants were usedfor the bioassays.

Insect

The eggs of Spodoptera liturawere collected from a field in TaichungCity, Taiwan and kept in 250-ml plastic rearing cups, and small wetcotton balls were added to provide moisture. The rearing cups were

Table 1Chemical properties of the field soil used in the study.

Property Value

pH 6.71EC (μS/cm) 30.9OM (%) 0.34Total N (g kg−1) 0.595P (mg kg−1) 76.13404K (mg kg−1) 35.75068Ca (mg kg−1) 443.2Mg (mg kg−1) 148.3595Fe (mg kg−1) 592.1538Mn (mg kg−1) 222.9418Cu (mg kg−1) 6.513184Zn (mg kg−1) 45.12512

stored in a growth chamber (25 °C, 16:8 h light/dark photoperiod;S.I.C. Co., Taiwan). After the eggs hatched, the larvae were also rearedin the 250-ml plastic rearing cups and fed an artificial diet (Yadavet al., 2010). Pupawere collected, separated according to sex, and placedin the 250-ml plastic rearing cups. After eclosion, the adults were paired(10 pairs) in a plastic cylinder (21 cm height × 14.9 cm diameter), andtissue paper was used to cover the inside for adults to oviposit eggs. Theplastic cylinders were placed under laboratory conditions and the adultmothswere fed a sugar solution (Yadav et al., 2010). The third instars ofthe insects were used for the bioassay.

Microbials and fertilizer treatments

To evaluate the effects of fertilization, microbials, and theirinteractions on plants and herbivores, three fertilization levels(zero, half, and full) and three groups of microorganisms (control,fungi, and bacteria) were used in this study. The fungi treatmentconsisted of only one fungi species, Meyerozyma guilliermondii.Two types of bacteria mixture were used for the bacterial treatment.The first type of bacterial mixture was labeled B1 and comprised threebacterial species: Burkholderia phytofirmans, Rhizobium miluonense, andRhizobium lusitanum. The second type of bacterialmixture (B2) containedtwo species: Bacillus subtilis and Ochrobactrum pseudogrignonense(Table 2). Each microbial used in this study was observed to improveplants’ ability to uptake nutrients and minerals (Hung et al., 2005;Nakayan et al., 2013; Rekha et al., 2007). Therefore, nine treatmentswere applied in this study: (1) no fertilization and no microbials,(2) half fertilization level and no microbials, (3) full fertilizationlevel and no microbials, (4) fungi only, (5) half fertilization leveland fungi, (6) full fertilization level and fungi, (7) bacterial mixtures,(8) half fertilization level and bacterial mixtures, and (9) full fertili-zation level and bacteria. Each microorganism used in this study wasprovided by the Laboratory of Soil Environmental Microbiology andBiochemistry, Department of Soil and Environmental Science, Na-tional Chung Hsing University. After the seeds were sown for 2weeks, the microbial suspension [F suspension (N108 cfu ml−1), B1suspension (N108 cfu ml−1), and B2 suspension (≥108 cfu ml−1)]were diluted 200 times using distilled water and then poured intoplastic trays. The seedlings in the 104-well plates were soaked inthe microbial suspension for 15 min as the first inoculation. After 2days, the seedlings were transplanted into 12.7-cm pots filled withfield soil. Every week after the transplant, 50 ml of various microbialsolutions were added into each pot, and 50 ml of water were addedto plants in the control treatments. Five microbial inoculationswere conducted in this study, and the concentration was increasedgradually to promote their effects. The microbials were diluted 100times using distilled water in the second and third inoculations,and were diluted 50 times using distilled water in the fourth andfifth inoculations (Table 3). In this study, we applied five times anddifferent concentration of PGPMs to the soil. The amount of PGPMswould decline through time; therefore, we would like to make surethat the soil contained the PGPMs. At the first week’s inoculation,the plants were small and only a low dose of PGPMs was applied.

Table 2The microorganisms used for the study.

Bacteria (B) B1a

Burkholderia phytofirmansRhizobium miluonenseRhizobium lusitanumB2a

Bacillus subtilisOchrobactrum pseudogrignonense

Fungi (F) Meyerozyma guilliermondii

a B1 and B2, mixture of the bacterial strains.

Table 3Microbial mixtures application process used for the PGPM treatments.

Inoculation Fungi Bacteria Concentration(PGPMs:water)

1st inoculation Fa B1b 1:2002nd inoculation F B2b 1:1003rd inoculation F B1 1:1004th inoculation F B2 1:505th inoculation F B1 1:50

a F, fungi.b B1 and B2, different bacterial mixtures.

589Y. Sripontan et al. / Journal of Asia-Pacific Entomology 17 (2014) 587–593

After the settle period, the plants grew well and had lots of roots, anincreased concentration of PGPMs was then used. Regarding thefertilizer treatments, a commercial synthetic fertilizer (20-20-20 N-P-K, Hyponex® 4; Hyponex Co., Marysville, OH, USA) wasused. The fertilizer was first dissolved in water and then added tothe soil. Plants were given 50 ml of water, half of the recommendedconcentration (0.5 g/1000 ml), or the recommended concentration(1 g/1000ml) for none, half-level, and full-level treatments, respectively.The fertilizer was treated weekly. The foliage of the cabbage (47 daysafter sowing) and tomato (51 days after sowing) plants were collectedfor plant performance analysis. The foliar water content, leaf area, andbiomass of aboveground parts were measured as indicators of plantgrowth performance.

Insect performance bioassay

The insect performance bioassay was conducted to evaluate theeffect of differently treated foliage on the relative growth rate ofS. litura. The seventh leaf of cabbage plants and the fourth leaf of tomatoplants were removed from the base of the plant by using surgicalscissors and placed individually into petri dishes (9 cm in diameter).The petioles of the leaves were kept in a 2-ml Eppendorftube with ROwater to maintain freshness. Third instar S. litura larvae were thenweighed and individually placed on differently treated leaves (25 °C,16:8 h light/dark photoperiod). The larvae were allowed to feed onthe foliage for 48 h. They were subsequently separated, frozen, oven-dried, and weighed. Ten replications (larvae) were conducted for eachtreatment. At the same timeof the bioassay, freshweights of 15 third in-star larvae were measured individually and then oven-dried at 45 °C.After 1 week, the dry weights of the larvae were measured again. Theaverage water content of the larvae was used to calculate the initiallarval dry weight used in the feeding study. The relative growth rate(RGR) was calculated using the following equation: ((final dry weightof insect − initial dry weight of insect) / initial dry weight of insect) /duration) (Farrar et al., 1989; Schoonhoven et al., 1998; Waldbauer,1968). RGR was used as an indicator of insect growth performance.

Foliar chemistry

In this study, we measured foliar protein content and polyphenoloxidase activity. The seventh leaf of the cabbage plant and the fourthleaf of the tomato plant were collected during the bioassay for foliarchemical analysis. Five plants were used for each plant species. Leafsamples were ground using liquid nitrogen and then homogenized ina 7% grinding buffer (polyvinylpolypyrrolidone in a potassium phos-phate buffer, pH 7). Leaf ground extract (1 ml) was mixed with 100 μlof 10% Triton X-100 (Sigma-Aldrich, St. Louis, MO, USA) in a microtube.The crude extract solution was then centrifuged at 4 °C at 10,000 rpmfor 15 min, after which the resulting supernatant was used for deter-mining enzyme activity. To quantify the amount of protein, a standardcurve was prepared using bovine serum albumin (BSA, Sigma-Aldrich,St. Louis, MO, USA) (Bradford, 1976; Tan et al., 2011, 2012).

Polyphenol oxidase activity was measured based on the proceduresof Stout et al. (1999) to calculate the formation rate of melanin-like

material from catechol. For this assay, 15 μl of a supernatant liquidwas mixed with catechol (0.1 M potassium phosphate buffer, pH 8).After mixing for 1 min, an absorbance value below 470 nmwas record-ed (Cipollini et al., 2004; Ryan et al., 1982; Thaler et al., 1996, 2001).

Statistical analysis

Mean and standard error valueswere calculated for plant growth,insect performance, and foliar protein and polyphenol oxidase content.Two factors of this study included PGPM and fertilizer. A two-wayanalysis of variance (ANOVA) and Tukey's multiple-range test (Version6.2; SAS Institute Inc., Cary, NC, USA, 1996)were used for comparing theinteraction effects between fertilization and microbial application.

Results

Plant growth performance

The effects of microbial application and fertilization on plant growthperformance (foliar fresh and dryweight and leaf area) differed. The re-sults indicated that the fresh weight of cabbage differed significantlyamong various microbial treatments (F = 35.98, P = 0.0001); fungitreatment exhibited the most significant effect on foliar fresh weight(Fig. 1). In addition, cabbage fresh weight increased markedly asthe level of fertilization increased (F = 560.33, P = 0.0001). Full-level fertilizer treatment produced a dry weight two times greaterthan that of the control treatment. Moreover, combined microbialand fertilizer application significantly affected the fresh weight ofthe cabbage (F = 4.94, P = 0.0028). The results also indicated thatthe foliar dry weight of the cabbage differed significantly betweenthe microbial (F = 18.76, P = 0.0001) and fertilizer (F = 87.80, P =0.0001) treatments. In addition, a significant interaction effect was ob-served betweenmicrobial and fertilizer application regarding the fo-liar dry weight (F = 8.00, P = 0.0001) (Fig. 1). Regarding the leafareas, both microbial (F = 47.53, P = 0.0001) and fertilizer (F =342.50, P = 0.0001) treatments exhibited profound effects. Fungitreatment using the full level of fertilization yielded the largest leafarea (Fig. 1). Additionally, combined microbial and fertilizer applica-tion significantly affected the leaf areas (F = 4.70, P = 0.0037).

The results indicated that the fresh weight of the tomato did notdiffer among the microbial treatments (F = 1.40, P = 0.2600). Bycontrast, nutrient availability significantly affected the fresh weight(F= 159.83, P= 0.0001). Full fertilization treatment doubled the fo-liar fresh weight (Fig. 2). Moreover, the effects of microbial and fertilizerapplication on foliar fresh weight were not significant (F = 1.22, P =0.3195). The dry weight of the tomato was significantly altered by bothmicrobial (F = 4.51, P = 0.0178) and fertilizer application (F = 84.78,P= 0.0001) (Fig. 2). However, the effects of microbial and fertilizer ap-plication on the foliar dry weight were independent of each other (nosignificant interaction) (F = 2.15, P = 0.0944). The results indicat-ed that the leaf area of the tomato was not affected by microbialtreatment (F = 2.92, P = 0.0670), whereas the level of fertilizationsignificantly influenced the leaf area (F = 151.19, P = 0.0001)(Fig. 2). In addition, the combined effect of microbial and fertilizerapplication on the leaf area was not significant (F = 0.33, P =0.8532).

Foliar chemistry

The effects of microbial and nutrient availability on nutrients(water and protein) and defense-related compounds (polyphenoloxidase, PPO) differed among treatments. Regarding the cabbage,foliar water content was observed to vary significantly among themicrobial treatments (F = 25.77, P = 0.0001), and overall, thefungi-treated cabbage exhibited greater water content (Fig. 3).In addition, fertilization slightly affected the foliar water content

Fig. 2. Growth performance of tomato plants treated with microbials and fertilizer.Mean ± SE (n = 5) (P b 0.05, Tukey’s test).

Fig. 1. Growth performance of cabbage plants treated with microbials and fertilizer.Mean ± SE (n = 5) (P b 0.05, Tukey’s test).


(F = 28.15, P = 0.0001). Similarly, a significant interaction effectalso occurred between microbial and fertilizer application regardingthe foliar water content (F= 10.59, P= 0.0001). The results indicatedthat protein content did not differ significantly among the microbialtreatments (F= 1.77, P= 0.1851), whereas fertilizer application sig-nificantly affected the protein content (F = 7.62, P = 0.0017)(Fig. 3). Moreover, the effect of microbial and fertilizer applicationon the cabbage’s protein content was nonsignificant (F = 1.51,P = 0.2200). Microbial treatment did not affect the PPO activity(F = 3.39, P = 0.0447), and the PPO activity also did not differsignificantly among fertilizer treatments (F = 3.86, P = 0.0302)(Fig. 3). However, the interaction between microbial and fertilizerapplication significantly influenced the PPO activity (F = 6.29,P = 0.0006).

Regarding the tomato, the results indicated that microbial applica-tion (F = 4.82, P = 0.0140) and the fertilization level (F = 26.74,P = 0.0001) significantly influenced the water content (Fig. 4).Moreover, the interaction between microbial and fertilizerapplication on the water content was not significant (F = 2.72,P = 0.0446). Protein content did not differ significantly amongthe microbial treatments (F = 0.61, P = 0.5515) (Fig. 4); however,

the fertilization level significantly affected the protein content (F =23.00, P= 0.0001). The fertilization treatment contained a higher pro-tein content than did the control treatment. However, the effect of thecombined microbial and fertilizer treatment on the protein contentwas nonsignificant (F = 1.02, P = 0.4093). The results revealedthat PPO activity was not affected by microbial application (F =4.64, P= 0.0161) (Fig. 4), whereas the fertilization level significant-ly affected the PPO activity (F = 11.76, P = 0.0001). Moreover, theeffect of microbial and fertilizer application on PPO activity also didnot differ significantly (F = 1.82, P = 0.1463).

Insect performance

The results indicated that microbial and fertilizer application pro-duced various effects on the performance of S. litura larvae. Regardingthe larvae that fed on the cabbage, the results indicated that microbial(F = 4.03, P = 0.0214) and fertilizer (F = 5.92, P = 0.0040)application significantly affected the RGR (Fig. 5). Moreover, the

Fig. 3. Chemical components of cabbage plants treated with microbials and fertilizer.Mean ± SE (n = 5) (P b 0.05, Tukey’s test).


interaction effect of microbial and fertilizer application on the perfor-mance of S. litura was significant (F = 4.31, P = 0.0033).

Regarding the larvae that fed on the tomato foliage, the results in-dicated that the RGR was not significantly influenced by microbialtreatment (F= 1.93, P= 0.1511) (Fig. 5). However, the fertilizationlevel significantly affected the performance of S. litura (F = 22.15,P = 0.0001). The larvae grew two times faster on the foliage whenusing the full fertilizer treatment than when using the control treat-ment. In addition, the interaction between microbial and fertilizer ap-plication on RGR was significant (F = 3.08, P = 0.0207).

Fig. 4. Chemical components of tomato plants treated with microbials and fertilizer.Mean ± SE (n = 5) (P b 0.05, Tukey’s test).

Discussion

The results of this study indicate that microbial application andfertilizer application produce significant and different effects on plantgrowth and insect performance. In addition, microbial treatment canpromote plant growth and significantly reduce the amount of fertilizerapplied.

Fertilizer may be the most crucial factor that affects plant growthand yield. Fertilizer application can promote plant growth. A previousstudy indicated that fertilized plants contain leaf numbers and plantbiomass five times higher than those of unfertilized plants (Hsu et al.,2009). However, high level of fertilization may also lead to high insectdamage levels. In addition to fertilizer, PGPMs in the soil environmenthave also been identified to play vital roles in plant growth. Studieshave revealed that several species of PGPMs, such as Bacillus sp. andBurkholderia vietnamiensis els., can significantly increase plant growthperformance in numerous plant species (Kokalis-Burelle et al., 2002;Rekha et al., 2007). Fertilizer and PGPMs may independently facilitateplant growth and a combined effect may occur when they are usedtogether. A previous study determined that combined PGPM and lowfertilizer application produces plant biomass and nutrient uptake levelssimilar to those of full fertilizer treatment (Adesemoye and Kloepper,2009; Afzal and Bano, 2008; Nakayan et al., 2013). Our results alsoindicated that microbials and fertilizer significantly influence plantgrowth. Moreover, we observed that plants treated with the fungi

Fig. 5. Relative growth rate of third instar larva of Spodoptera litura on cabbage plants(A) and tomato plants (B) treated with microbials and fertilizer. Mean ± SE (n= 10)(P b 0.05, Tukey’s test).


Meyerozyma guilliermondii and a half level of fertilization generate aplant biomass similar to that generated when using full fertilizertreatment, suggesting that adding these microbials can reduce theamount of fertilizer required and, consequently, reduce farming costs.

Fertilizer and PGPM application may affect plants’ chemistry (Hsuet al., 2009; Lucy et al., 2004; Peric et al., 2009; Stout et al., 1998).Fertilizer normally exhibits a significant influence on the primaryplant metabolites. Previous studies have indicated that high nutrientavailability increases foliar protein and nutrient content (Bybordi andEbrahimian, 2013; Peric et al., 2009; Prudic et al., 2005; Stout et al.,1998). The results of the current study also suggested that full fertilizertreatment produces twice the protein content. In addition, the levels ofsecondary plant metabolites, such as defensive proteins (trypsin inhib-itor (TI) and PPO) and phenolics, can also be elevated by fertilization;this maybe due to an increase in resource availability (Hsu et al.,2009; Peric et al., 2009; Stout et al., 1998). The PPO activity of thetomato plant, however, decreased when applying fertilizer in thisstudy, and the cause remains unclear. Moreover, because of theircharacteristic of enhancing root absorbing efficiency, PGPMs havebeen considered to influence the nutrient uptake of host plants and toincrease the macro- and micronutrient and protein content of plants(Bashan and de-Bashan, 2005; Egamberdiyeva and Höflich, 2003;Glick, 2004; Lai et al., 2008). PGPMs have been observed to increasethe nitrogen content and mineral content of various plant species(Belimov and Dietz, 2000; Boddey and Dobereiner, 1988; Cakmakciet al., 2001; Dobbelaere et al., 2001; Probanza et al., 2002). Similarly,relevant literature has also indicated that PGPMs induce the productionof certain secondary plant metabolites (Figueiredo et al., 2011;Shanmugam and Kanoujia, 2011). Previous studies have determinedthat plants treated with PGPR increase the activity of peroxidase, PPO,phenylalanine ammonia-lyase, and proteinase inhibitors (Chen et al.,2000; Figueiredo et al., 2011; Shanmugam and Kanoujia, 2011).

However, little is known about the interaction between the two factors;only a few studies have indicated that the effect of PGPMs and fertilizerincreases the concentrations of macronutrients (N, P, K, and Ca) andmicronutrient (Mg, Fe, Zn, Cu, and Mn) in plants (Lai et al., 2008;Nakayan et al., 2013). Our results suggest that combined microbialand fertilizer treatment significantly increases foliar protein content.However, the PPO activity decreased during combined treatment.Therefore, the effect of PGPMs and fertilizer on the nutrient content ofplants can be confirmed; however, the effects on plant defensivecompounds remain unverified. Additional studies are required toclarify the effect and the mechanisms of PGPMs on the nutrientallocation of plants.

Fertilizer and PGPMs are two vital factors that influence a plant’schemistry and subsequently affect herbivorous insects’ performance(Herms, 2002; Pineda et al., 2010; Ramamoorthy et al., 2001). Previousstudies have indicated that fertilized plants produce small amounts ofdefensive chemicals; thus, herbivorous insects perform better onfertilized plants than on unfertilized plants (Altieri and Nicholls, 2003;Hsu et al., 2009; Prudic et al., 2005). PGPMs promote plant growth by in-creasing root absorption; however, relevant literature has reported thatPGPMsmay also induce a plant’s defense responses (Pineda et al., 2010;Ramamoorthy et al., 2001). For example, a previous study revealed thatPGPR-treated plants are resistant to feeding by the cucumber beetleDiabrotica undecimpunctata howardi Barber (Zehnder et al., 1997,2001), leaf folder Cnaphalocrocis medinalis (Saravanakumar et al.,2007), and leaf miner Aproaerema modicella (Senthilraja et al., 2010).Fertilization and PGPMs exhibit varied effects on plants and,consequently, on herbivorous insects; however, the combined PGPRand fertilizer effect on insect performance is largely unknown. Ourresults indicate that microbial application and fertilization couldsignificantly affect the RGR of the third instar larvae of S. litura; butthe interaction effect is still not clear.

In summary, our study suggests that PGPMs and fertilizer exertvarious influences on plant growth performance, foliar chemistry, andinsect performance. Both treatments exhibit positive effects on plantperformance. However, the interaction effect of the microbials andfertilizer on plants remains unclear. Therefore, additional studies arerequired to clarify the interaction effect when using them together inpromoting crop production and pest control.

Acknowledgments

We thankWallace Academic Editing and two anonymous reviewersfor their comments on the manuscript.

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Effects of plant-growth-promoting microorganisms and fertilizers on growth of cabbage and tomato and Spodoptera litura pe...IntroductionMaterials and methodsPlantsInsectMicrobials and fertilizer treatmentsInsect performance bioassayFoliar chemistryStatistical analysis

ResultsPlant growth performanceFoliar chemistryInsect performance

DiscussionAcknowledgmentsReferences

Journal of Asia-Paci c Entomology · Effects of plant-growth-promoting microorganisms and...

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Transcript of Journal of Asia-Paci c Entomology · Effects of plant-growth-promoting microorganisms and...