The results of ourwork show that th

The results of our
work show that there are some antagonist yeasts among the
microbial community associated with grape berries which are able
to control black aspergilli. To our knowledge, this is the first report
in which yeasts have been isolated in Cyprus and assessed for
potential antagonistic ability against A. tubingensis, the most frequently
isolated Aspergillus species from Cyprus vineyards (unpublished
data). Traditionally, yeasts identification relied on
morphological, physiological and biochemical characteristics.
These classical microbiology methods are laborious and time consuming
and exhibit restricted sensitivity and specificity. As a consequence
traditional methods are being successfully replaced by
several molecular biology approaches. The molecular methods
are based on DNA fingerprinting by means of RAPD-PCR, arbitrarily
primed PCR (AP-PCR) and fluorescent AFLP (fAFLP) (De Curtis et al.,
2004; Schena et al., 1999, 2000, 2002) and on polymorphisms and
sequence analysis of DNA regions encoding the rRNA genes
(Kurtzman and Robnett, 1998) or the non-coding ITS (Internal
Transcribed Spacers) (Cadez et al., 2002; Sabate et al., 2002) and
IGS (Intergenic Spacer) regions (Diaz and Fell, 2000). The experimental
data presented in this paper demonstrate that the epiphytic
population identified belonged to seven taxonomically distinct
groups (Table 1) based on sequencing of the D2 region of the
nuclear LSU rRNA. A total of 55 identified yeast isolates were evaluated
for their antagonistic effect against A. tubingensis by an
in vitro co-inoculation assay performed on agar plates and it was
shown that 33 isolates (60% of the analyzed population) were able
to inhibit fungal sporulation (Table 2 and Fig. 1). Although fungal
growth in dual cultures was not severely affected, the antisporulant
effect was considered as indicative of fungal sensitivity to
the action of yeasts isolated from the same biological niche.
Indeed, in the detached berry test 28 isolates, representing 82%
of the analyzed population (previously shown to inhibit fungal
sporulation in vitro) showed significant biocontrol activity against
A. tubingensis by reducing its ability to colonize wound inoculated
berries (Table 3 and Fig. 2). Significant differences were observed
among assayed yeasts in terms of% inhibitory activity, with A. pullulans
being the most, and C. sake and C. zeylanoides the least effective
in inhibiting A. tubingensis growth (Table 3). Our findings
showed that biocontrol efficacy is a strain-related characteristic
and is not depended only on species or genus, as observed in previous
studies (Bleve et al., 2006; Suzzi et al., 1995).
The screening of epiphytic yeasts in the present study identified
two isolates of C. magnus (2ZK2 and XM19) and one isolate of C.
sake (2AM3) that reduced A. tubingensis sporulation on agar plates
as well as fungal growth on grape berries. These findings are consistent
with those of previous studies in which various isolates and
strains of Candida spp. were reported to be effective against several
fungal pathogens (Bleve et al., 2006; Guinebretiere et al., 2000;
Lima et al., 1997; McLaughlin et al., 1990, 1992; Vinas et al.,
1988; Zahavi et al., 2000). Moreover, the results of this study
revealed that A. pullulans was both the predominant yeast species
on grapes (37 out of 55 isolates) and the most effective biocontrol
agent, showing the highest degree of antagonistic activity against
A. tubingensis. More specifically, 73% of the isolates reduced A. tubingensis
sporulation in agar plates and 92.6% inhibited colonization
of grape berries by the same fungus. Strains of A. pullulans were
previously reported to be effective biological control agents
(BCAs) exhibiting antagonism to several pathogens on different
crops, including: (a) Botrytis cinerea, Penicillium expansum,
Rhizopus stolonifer and A. niger on table grapes (Castoria et al.,
2001), (b) the OTA producer A. carbonarius on table grapes and
strawberry (Castoria et al., 2001; De Curtis et al., 2012b; de
Felice et al., 2008; Lima et al., 1997), (c) B. cinerea and P. expansum
on apples as well as B. cinerea and Monilinia laxa on sweet cherries
and table grapes (Bencheqroun et al., 2007; Castoria et al., 2001;
Ippolito et al., 2000, 2005; Janisiewicz et al., 2000; Leibinger

ผลลัพธ์ (ไทย) 1:

ผลการใช้ของเราทำงานมียีสต์ปฏิปักษ์บางหมู่ชุมชนจุลินทรีย์ที่เกี่ยวข้องกับองุ่นผลเบอร์รี่ซึ่งจะสามารถการควบคุม aspergilli สีดำ ความรู้ของเรา เป็นรายแรกซึ่งยีสต์มีการแยกประเทศไซปรัส และประเมินศักยภาพความสามารถในการเป็นปรปักษ์กับ A. tubingensis บ่อยที่สุดสายพันธุ์ Aspergillus แยกจากไร่องุ่นไซปรัส (ยกเลิกประกาศข้อมูล) ประเพณี พึ่งรหัสยีสต์ลักษณะสัณฐาน สรีรวิทยา และชีวเคมีวิธีการทางจุลชีววิทยาคลาสสิกเหล่านี้จะลำบาก และใช้เวลานานและจำกัดความไวและความจำเพาะ เป็นผลวิธีการแบบดั้งเดิมจะถูกแทนเรียบร้อยโดยวิธีการชีววิทยาโมเลกุลต่าง ๆ วิธีการระดับโมเลกุลขึ้นอยู่กับดีเอ็นเอลายพิมพ์ โดยเทคนิค RAPD โดยพลการงเยีย PCR (AP-PCR) และ AFLP เรืองแสง (fAFLP) (เดเคอร์ติส et al.,2004 Schena et al. 1999, 2000, 2002) และครั้งที่สอง และวิเคราะห์ลำดับของดีเอ็นเอเข้ารหัสยีน rRNA(Kurtzman และ Robnett, 1998) หรือไม่ใช่รหัสของ (ภายในทับศัพท์ในรูปแหวน) (Cadez et al. 2002 Sabate et al. 2002) และIGS (Intergenic Spacer) ภูมิภาค (ดิแอซและล้มลง 2000) การทดลองข้อมูลที่นำเสนอในเอกสารนี้แสดงให้เห็นว่าที่บนต้นไม้ระบุว่าประชากรเป็นเจ็ดแตกต่างจัดหมวดหมู่กลุ่ม (ตารางที่ 1) ตามลำดับของภูมิภาค D2 ของการnuclear LSU rRNA. A total of 55 identified yeast isolates were evaluatedfor their antagonistic effect against A. tubingensis by anin vitro co-inoculation assay performed on agar plates and it wasshown that 33 isolates (60% of the analyzed population) were ableto inhibit fungal sporulation (Table 2 and Fig. 1). Although fungalgrowth in dual cultures was not severely affected, the antisporulanteffect was considered as indicative of fungal sensitivity tothe action of yeasts isolated from the same biological niche.Indeed, in the detached berry test 28 isolates, representing 82%of the analyzed population (previously shown to inhibit fungalsporulation in vitro) showed significant biocontrol activity againstA. tubingensis by reducing its ability to colonize wound inoculatedberries (Table 3 and Fig. 2). Significant differences were observedamong assayed yeasts in terms of% inhibitory activity, with A. pullulansbeing the most, and C. sake and C. zeylanoides the least effectivein inhibiting A. tubingensis growth (Table 3). Our findingsshowed that biocontrol efficacy is a strain-related characteristicand is not depended only on species or genus, as observed in previousstudies (Bleve et al., 2006; Suzzi et al., 1995).The screening of epiphytic yeasts in the present study identifiedtwo isolates of C. magnus (2ZK2 and XM19) and one isolate of C.sake (2AM3) that reduced A. tubingensis sporulation on agar platesas well as fungal growth on grape berries. These findings are consistentwith those of previous studies in which various isolates andstrains of Candida spp. were reported to be effective against severalfungal pathogens (Bleve et al., 2006; Guinebretiere et al., 2000;Lima et al., 1997; McLaughlin et al., 1990, 1992; Vinas et al.,1988; Zahavi et al., 2000). Moreover, the results of this studyrevealed that A. pullulans was both the predominant yeast specieson grapes (37 out of 55 isolates) and the most effective biocontrolagent, showing the highest degree of antagonistic activity againstA. tubingensis. More specifically, 73% of the isolates reduced A. tubingensissporulation in agar plates and 92.6% inhibited colonizationof grape berries by the same fungus. Strains of A. pullulans werepreviously reported to be effective biological control agents(BCAs) exhibiting antagonism to several pathogens on differentcrops, including: (a) Botrytis cinerea, Penicillium expansum,Rhizopus stolonifer and A. niger on table grapes (Castoria et al.,2001), (b) the OTA producer A. carbonarius on table grapes andstrawberry (Castoria et al., 2001; De Curtis et al., 2012b; deFelice et al., 2008; Lima et al., 1997), (c) B. cinerea and P. expansumon apples as well as B. cinerea and Monilinia laxa on sweet cherriesand table grapes (Bencheqroun et al., 2007; Castoria et al., 2001;Ippolito et al., 2000, 2005; Janisiewicz et al., 2000; Leibinger