Assessment of Bacterial Blight Tolerance of Persian Walnut Based on Immature Nut Test

Transcription

1 Avalle Band A onlne: et al. / Not Sc Bol, 2015, 7(2): Prnt ISSN ; Electronc Not Sc Bol, 2015, 7(2): DOI: /nsb Assessment of Bacteral Blght Tolerance of Persan Walnut Based on Immature Nut Test Attla BANDI 1, 2 *, Mára HEVESI 1, Zsolt SZANI 3, Magdolna TÓTH 1 1 Corvnus Unversty of Budapest, Faculty of Hortcultural Scence, Department of Pomology, Vllány str , Budapest, Hungary; attla.band@un-corvnus.hu; mara.heves@un-corvnus.hu; magdolna.toth@un-corvnus.hu 2 Sapenta Unversty, Faculty of Techncal and Human Scence, Department of Hortculture, Sghșoara str. 1/C, Tîrgu-Mureș, Romana; bandattla@ms.sapenta.ro (*correspondng author) 3 Natonal Food Chan Safety Offce, Varety Testng Department for Hortcultural Crops, Kelet K. str. 24, Budapest, Hungary; SzanZs@nebh.gov.hu Abstract Bacteral blght s one of the most serous dseases affectng Persan walnut (Juglans rega L.). Susceptblty to ths dsease was evaluated by artfcal noculaton n an n vtro experment for cultvars developed n Hungary and for s from Transylvana. Thrty peces of mmature frut of 11 walnut cultvars and 10 s were noculated by puncton usng a suspenson contanng a mxture of Xaj-solates of controlled vrulence. As control, a moderate resstant (mr) Pedro and a hghly susceptble (hs) Mlota ntenzív cultvars were used. After ten days the dameter of the necrotc area around the noculaton ponts was measured and the dsease rate (DR) was noted. For the calculaton of ndexes t was taken the structure of the lesons (dffuse or defned margn) nto consderaton. None of the 21 cultvars/s analysed were found to be hardly resstant, although all of them showed a hgher degree of resstance than the susceptble control ( Mlota ntenzív ). The majorty of the analysed cultvars ft nto the moderate susceptble (ms) and moderate resstant (mr) group. Mlota kése and SZEN-10, had a notle reacton as consdered to be moderate resstant (mr), showed a smlar degree of resstance compared to the control cultvar Pedro. It was concluded that these cultvars, based on ther resstance to Xanthomonas arborcola pv. juglands, could be proposed as resstance gene sources, as well as for producton purposes n the envronmental condtons of the Carpathan Basn. Keywords: artfcal noculaton, n vtro, solates, Juglans rega L, stress, Xanthomonas Introducton Walnut trees may suffer stress from otc or botc envronmental factors, whch can lead to crop falure. The bacteral blght dsease, Xaj (Xanthomonas arborcola pv. juglands (Perce)) (Vautern et al.,1995) s one of the most serous dseases affectng Persan (Englsh) walnut (Juglans rega L.) (Loret et al., 2001). The dsease has been known snce the end of the 19 th century (Ferrars, 1927) and s wdespread n walnut growng areas. It causes severe damage to leaves, twgs, buds, petoles, rachdes, male and female catkns, nutlets and kernels, and s consdered to be a major cause of reducton n frut yeld and tree vgour (Belsaro et al., 1999). The damage produced by ths pathogen s predsposed by wet sprngs. Rany sprngs, dew and contnual hgh humdty condtons are favourle to the development of severe blght, resultng n sgnfcant crop loss (Belsaro, 1997). If ths happens just before and after the flowerng tme, t may cause losses of 50% to 80% of the crop (Charlot and Radx, 1993). The bactera enter the walnut tssues through natural openngs, such as stomata and damaged tssues (Sharma and Sharma, 1999). All succulent new growths are reported to be very susceptble to the dsease, becomng more resstant when maturng (Belsaro et al., 1997). Susceptble cultvars may have an ncreased level of damage on new plantngs. Soon after the walnut blght was frst descrbed (Perce, 1901), there were many reports of resstance to the dsease (Soltan and Alad, 2010). Resstance of trees has been evaluated n orchard condton, so t has not been clarfed whether the sence of blght was due to dsease escape or to an unknown form of resstance. A dfferent susceptblty to walnut blght wthn walnut cultvars has been reported. Several authors, Aletà and Nnot (1993), German (1997), Woeste et al. (1992) have dscovered no resstant cultvars; however Franquette and Hartley were reported as resstant cultvars (Belsaro, 1997; Belsaro et al., 1999). Usng the n vtro method, assessment of the resstance of walnut genotypes to bacteral blght has been carred out n several studes (Belsaro, 1997; Belsaro et al., 1999; Martns, 1997; Soltan and Alad, 2010; Özaktan et al., 2007; Tsantos et al., 2008). Detaled cultvar examnaton offers the potental of reducng the dsadvantageous nfluence of the man stress factors (Szan, 2009). Searchng for the sources of resstance s mportant, snce large walnut trees are not easy to treat, and furthermore, ncreasngly copper-resstant Xaj strans are developng (Solar et al., 2007). Receved: 09 June Receved n revsed form: 17 June Accepted: 17 June Publshed onlne: 21 June 2015.

2 254 Tle 1. Examne ed walnut cultvars and s Cultvar or denomnaton Hartley Pedro Mlota 10 Alsószentván 117 Alsószentván 118 Mlota ntenzív Mlota kése Mlota bőtermőő Alsószentván kése Bonfác M SZEN-10 ALB-22 FFA-11 OZSD-37 SAR-33 SOM-50 SOM-90 SOM -101 SOM-120 SOM-274 a Tulecke and McGranahan, 1994 b Szentvány, 1990 Band A et al. / Not Sc Bol, 2015, 7(2): Parentage Franquette x Mayette a Conway Mayette x Payne a Selecton Selecton Selecton Mlota 10 x Pedro b Mlota 10 x Pedro b Mlota 10 x Pedro b Alsószentván 117 x Pedro b Alsószentván 117 x Pedro b Mlota 10 x Pedro b Orgn USA USA Tle 2. The evalu uaton of dsease severty based on the nfecton of mmature walnut fruts Dsease appearance/dameter of spots (mm) No symptoms Less than 1.00 mm, superfcal and dffuse spots on the noculaton pont Slght blackenng on the noculaton pont of nut by 1.01 mm to 2.00 mm Blackenng on the noculaton pont of nut by 2.01 mm to 5.00 mm of delved spot Blackenng on the noculaton pont of nut more than 5.01 mm of crater- type spot Scale of susceptblty Based on the results of prevous evalua atons and open feld examnatons (Thesz et al., 2007, 2009; Szan, 2009), ther ere was dentfed a dfferent degree e of susceptbl lty/resstance n the analysed popul ulatons and regstered cultvar rs. However, n most open feld asses essments, the dsease was detec cted by means of the leaf. The degre ee of frut nfect ton has been nvestgated to a lesser extent. Theref fore, t was consdered necessary to carr ry out artfcal nocul laton n contro olled crcumsta ances. The am of the study was to evaluate the susceptblty to Xaj of the cultvars mproved n Hungary, and of the ns from Transyl ylvana, by means of an artfcal noculaton techn nque n an n vtro experment. Materals and methods Plant mate eral Durng the research betw ween 2010 to 2013, t was evaluated the degree of susceptblty of the cultvars lsted n Tle 1, n comparson to Pedro, moderate resstant t (mr) and Mlota ntenzív hghl ly susceptble (hs), used as control cultvars s. The samples were collected before e shell hardenn ng n June, from the Tral Staton of the Naton nal Food Chan Safety Offce at Pölöske () and from farm mer orchards n Transylvana (). Bactera For the artfcal noculat ton there were e collected 61 solates from dfferen nt walnut-gro owng areas of Hungary and Transylvana. The lty to nduce hyperse enstve reacto on was examned on the tobacco leaf - Ncotana tacum L. (Kle ement, 1963) - and bean pod (Phase eolus vulgars L..). The pathoge encty Fg. 1. The results of pathogencty test on mmature walnut frut (A: SOM-120; B: ALB-22) test was controlled on mmature walnut fruts (Fg. 1). For the artfc cal noculaton n, t was used a mxture of solates from a homogeneous bactera populaton on (based on the phenotype e) from the green husk of two walnu ut fruts, one orgnatng from Hung ngary B02489 (), and the other from Transylvan a B (). They showed the same degree of vrulence. The boch hemcal dent fcaton of the two solate es (henceforth h, stran n ) was already carred out n prevous stud es (Band et al., ). Inoculaton For the artfcal al noculaton t was adjusted a suspenson of 10 8 cell.ml -1 concen ntraton utlsn ng a spectroph hotometer (570 nm wavelength) from the respect tve strans. The susceptblty ty

3 Band A et al. / Not Sc Bol, 2015, 7(2): Mean dameter (mm) SAR-33 SOM-274 Mlota ntenzív OZSD-37 ALB-22 SOM-90 SOM-50 FFA-11 Mlota 10 Bonfác Alsószentván 117 SOM-120 Alsószentván 118 Alsószentván kése Hartley Pedro M Mlota bőtermő Mlota kése SOM SZEN-10 j j g g h h a bc cd cd de ef f Mean dameter (mm) Fg. 2. Mean dameter of spots produced by two Xaj solates around the noculaton ( ) Values wth the dfferent letter ndcate cultvars that dffer sgnfcantly (Duncan s test, p 0.05); The error bars show SE of the mean tests were conducted based on the method of Özaktan et al. (2008). Durng the nvestgaton, 30 crops (20 for artfcal noculaton and 10 for control treatment) were collected. Each of the 20 frut was njected n fve (0.5 cm 2 ) places wth a 20 μl bacteral suspenson. The susceptblty of each cultvar was evaluated for 100 nstances of nfecton. The control fruts were noculated wth sterle dstlled water. The noculated fruts were then placed n closed transparent plastc boxes n order to assure an adequate humdty (85%) and temperature (26-28 C). Assessment of the susceptblty of fruts After noculaton, the fruts were montored contnuously, and on the tenth day the symptoms were summarsed. In order to determne the mean dameter of the necrotc area, a dgtal slde-callper attached to a computer (provdng accuracy to the mllmetre) was used. To determne the degree of the susceptblty, there were taken nto consderaton all the developed symptoms, as well as those that dd not appear. After ths, the dmensons of the necrotc areas were evaluated based on the fve-stage scale developed by Özaktan et al. (2008) (Tle 2). Determnaton of cultvar susceptblty Usng data from the nfecton scale and the formulas found n the related lterature, dfferent ndexes were produced. The dsease ratng (as an ndex showng the ncdence of the dsease) was determned by usng data from the nfecton scale and the followng formula by Bertrand and Gottwald (1986): Dsease rate= (a xf )/n where a - scale value (ndex of nfecton), f - frequency referrng to the dsease rate; n- the number of analysed frut per cultvar. Statstcal evaluaton The data were evaluated usng the SPSS software (IBM SPSS 22.0, Chcago, IL). The statstcal analyses were determned Fg. 3. Dendogram based on the average lnkage (between groups) method of cluster analyss by means of necrotc spot dameter and dsease rate of the walnut blght nfecton for walnut cultvars ( ) Numbers on the dendogram ndcate dfferent groups: 1: moderate resstant (mr), 2: moderate susceptble (ms), 3: susceptble (S), 4: hghly susceptble (hs) based on the sample sze, dstrbuton analyss (Kolmogorov- Smrnov test) and equvalence of devaton (Levene test). Where the condton of normalty was fulflled, and means proved to be homogeneous, Tukey s multple range test was used. If homogenety of dstrbuton was not fulflled, dfferences were analysed usng the Games-Howell test. In case of an normal dstrbuton, the Kruskal-Walls non-parametrc method was appled; then, for the comparson of the pars, the Mann- Whtney test was used. It was accepted the p 95% confdence level. The results were gven as average ± standard error (SE). It was conducted a herarchcal cluster analyss based on four-year data regardng the dameter and dsease rate, n order to classfy the cultvars nto susceptblty groups. The results were represented on a dendogram. Results and dscussons The susceptblty of the analysed cultvars was evaluated n dfferent groups accordng to the expermental year. Tle 3 contans annual mean values regardng the dsease rate. In the case of several cultvars, major dfferences between the annual results were noted. Therefore, further research wll be requred to clarfy the susceptblty of Alsószentván 118 and Mlota kése. Based on the results of the artfcal noculaton from 2010, compared to other cultvars, the mean dsease rate value of SZEN-10 was sgnfcantly lower (p<0.05), whch means t can be desgnated to the moderate susceptble class. OZSD- 37, SOM-90, ALB-22 and SOM-50 cultvars proved to be hghly susceptble. In the case of data from 2011, the Mann- Whtney test classfed the cultvars wth the average dsease rate value under 2 nto the homogeneous group (moderate susceptble), beng sgnfcantly dfferent (p 0.05). Consderng the 2012 dsease rate results, the susceptblty of cultvars showed a better dfferentaton. Based on the statstcal evaluaton of data, SOM-101 and SZEN-10 proved

4 256 Tle 3. Dsease rate of the analysed walnut cultvars n the ndvdual years a Band A et al. / Not Sc Bol, 2015, 7(2): Cultvar or Dsease rate (0-4) 2010 b 2011 c 2012 b 2013 c Hartley 2.91±0.03 a-c 2.19±0.04 g Pedro 1.64±0.21 gh 2.10±0.03 gh Mlota ±0.24 de 2.90±0.05 a-d Alsószentván ±0.16 d-f 2.64±0.05 de 2.88±0.10 d Alsószentván ±0.38 gh 2.53±0.02 ef Mlota ntenzív 2.55±0.20 a-c 2.57±0.24 b-e 2.80±0.08 b-e Mlota bőtermő 2.31±0.06 ef 2.18±0.08 g Mlota kése 1.29±0.29 h 2.16±0.01 g M ±0.04 gh 2.21±0.01 fg Alsószentván kése 2.72±0.03 a-e 2.21±0.05 fg Bonfác 2.13±0.40 bc 2.50±0.08 c-e 2.95±0.01 a-d ALB ±0.08 a 3.09±0.03 a 2.96±0.02 a-d 3.06±0.00 a FFA ±0.12 a-c 2.95±0.03 a-c 2.93±0.03 a-d 3.00±0.00 bc OZSD ±0.01 a 3.08±0.01 a 2.98±0.03 a-c 3.01±0.01 b SAR ± ±0.23 a-c 3.06± ±0.05 a SZEN ±0.10 c 2.24±0.09 fg 1.89±0.13 gh 2.48±0.21 e SOM ±0.06 a 2.91±0.08 a-c 3.10± ±0.01 c SOM ±0.01 a 3.00± ±0.06 a 3.00±0.00 bc SOM ±0.05 a-c 1.61±0.09 gh 1.81±0.09 h 2.30±0.03 f SOM ±0.12 a-c 2.79±0.06 a-d 2.69±0.06 c-e 2.88±0.03 d SOM ±0.09 a-c 3.00± ±0.11 a-d 3.00±0.00 bc a The mean values±se of the cultvars n a gven year b The dfferent letters ndcate the sgnfcant dfference between the cultvars (p 0.05; Tukey s multple range test) c The dfferent letters ndcate the sgnfcant dfference between the cultvars (p 0.05; Mann-Whtney test) to be moderate resstant (mr), whle moderate susceptblty (ms) was detected for Pedro, Mlota kése, Mlota bőtermő, Hartley, M and Alsószentván kése cultvars. Based on the average data from 2013, smlar to the results of prevous years, t was determned a sgnfcantly low susceptblty n the case of SOM-101 and SZEN-10. Fg. 2 contans the mean data for the dameter of the necrotc spots. Mlota ntenzív, ncluded n the experment as the susceptble control, proved to be the thrd most susceptble. Smlar, hghly susceptble cultvars were SAR-33, SOM-274, OZSD-37 and ALB-22. Compared to the moderate resstant (mr) Pedro control cultvar, SZEN-10 and SOM-101 showed a major resstance and these dfferences were statstcally sgnfcant (p< 0.05). The dendogram (Fg. 3), contanng the annual dameter of the necrotc spots and the dsease rate values, dfferentated the cultvars belongng to certan susceptblty classes from the susceptble group to less susceptble ones. The results from dfferent years regardng the degree of susceptblty/resstance of cultvars showed a correspondence for the majorty of cultvars; n the case of those cultvars, whch showed a major dfference between the annual results, t was accepted the classfcaton from the dendogram. Rovra et al. (2007) consdered the artfcal noculaton appled to be mportant because open feld recordngs of cultvar susceptblty are not suffcent. The reacton of the studed cultvars to the Xaj bactera was measured for the frst tme usng n vtro experment wth an noculaton method; therefore, only the control cultvar data can be compared to prevous results. Several studes provde nformaton regardng the degree of susceptblty to bacteral blght of Hartley and Pedro. Özaktan et al. (2007, 2008) evaluated the susceptblty of several cultvars wth artfcal noculaton, and based on data from 2007, t was determned a low susceptblty for Pedro (blght severty=14.1%) and a moderate susceptblty accordng to data from 2008 (blght severty=55%). In the current expermental data, n 2011 Pedro showed a moderate resstance, whle n 2012 t was determned moderate susceptble. Informaton out the susceptblty of Hartley and Mlota 10 cultvars can be obtaned from the results of Tsantos et al. (2008). Based on the 2007 and 2008 Varetal Susceptblty Index (VSI) values and the susceptblty classes, Hartley proved to be moderate susceptble (VSI = 25; Class of Susceptblty=B), whle Mlota 10 was found susceptble (VSI = 80, Class of Susceptblty = D). Solar et al. (2008, 2009) also determned a moderate susceptblty n case of Hartley cultvar. Accordng to the current results Hartley turned out to be susceptble n 2011, yet moderate susceptble n Thesz et al. (2007) consdered the Transylvanan cultvars to have a low susceptblty aganst Xaj. Accordng to the seven year results of Szan (2009), cultvars mproved n Hungary showed a low or moderate nfecton rate. Based on the annual measurements t can be concluded that the majorty of analysed cultvars were moderate susceptble (ms) and susceptble (S). Among the cultvars mproved n Hungary, Mlota kése, Alsószentván 118, M and Mlota bőtermő showed a low degree of susceptblty. Among the Transylvanan s, SZEN-10 and SOM- 101 showed a notle reacton, snce accordng to the annual results they were determned wth moderate resstance (mr) and a moderate susceptblty (ms). Conclusons The present research provded comprehensve results regardng the degree of susceptblty/resstance of the studed cultvars. Prevous open feld Xaj assessments based on the leaf qualty proved that the cultvars have a good resstance to the dsease. However, durng the current experment regardng artfcal noculaton of mmature fruts, t was

5 Band A et al. / Not Sc Bol, 2015, 7(2): proved that several cultvars have Xaj susceptblty. Therefore, the evaluaton based on the leaf was found not relle. Thus t may be concluded that the Pedro transmtted the Xaj resstance nto certan progeny. Ths could prove effectve as a gene source n further mprovements and for producton n the envronmental condtons of the Carpathan Basn. The results of these analyses show that, wth adequate results regardng cultvar susceptblty, the losses caused by Xaj bactera can be reduced. Acknowledgements Ths work was supported by the TAMOP /B- 09/1/KMR project. References Aletà N, Nnot A (1993). Varedades de nogal. Frutcultura Profesonal 54: Band A, Tóth M, Heves M (2014). Comparson of Xanthomonas arborcola pv. juglands solates from walnut trees grown n Romana and Hungary. Internatonal Journal of Hortcultural Scence 20(1-2): Belsaro A (1997). Susceptblty and factors predsposng to Xanthomonas campestrs pv. juglands n walnut nurseres. 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