University of Agriculture Faisalabad - Dr. Abdul Rehman Assistant … · 2012-04-14 · Dr. Abdul...
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Dr. Abdul RehmanAssistant Professor, Department of plant pathology, University of Agriculture Faisalabad.
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To study the Morphological characterization of Ceratocystis manginecans sp. nov. isolates from Pakistan
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In our country export quality mango fruit is produced but unfortunately we are facing with the low production of mango due to the mango sudden death syndrome disease.
Symptoms on trees include gum exudation with severe rotening smell, bark splitting, vascular discoloration with black and white streaks, total or partial wilting of the tree by succeeding browning of
leaves small holes with galleries due to the presence of bark beetle
(Hypochryphalus mangifera Stebbing: Coleoptera Scolytidae) (Al-Adawi 2002; Al-Adawi et. al., 2003).
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The fungal isolates recovered from affected mango trees wooden tissues in Pakistan shows distinct as Ceratocystis manginecans from species complex of C. fimbariata sensu stricto and sensu lato.
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Sampling:Several mango orchards were visited from Khanewal, Multan, Muzzaffergarh and Rahim Yar Khan during 2010 to 2011. Stem white and black streaked cross sections were collected from the affected mango trees.
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Isolation was done by the carrot method (Moller and De Vay, 1968)
Ascospore from ascomatal masses were collected and purified on 2% malt extract agar media (MEA).
All isolates are maintained in Disease diagnostic lab, Department of Plant pathology, University of Agriculture Faisalabad.
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Morphological characterization of isolates was done as, The ascomatal bases were globosed,
black (153- ) 192-254 (-281) µm in diameter. Ascomatal necks were dark brown to black becoming lighter at apices
(514- ) 557-635 (-673) µm long, (25- ) 32-42 (-48) µm wide at base, (14- ) 16-22 (-26) µm wide at apices.
Ostiolar hyphae were hyaline and divergent (42- ) 45-59 (-69) µm long.
Ascospores were hyaline, hat-shaped (3-4) µm long, (4-5) µm wide excluding sheath and (7-8) µm wide including sheath.
(Van Wyk, et al., 2007)
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Primary cylindrical-shaped hyaline conidia were long (15- ) 23-29 (-33) µm wide (3- ) 4-6 (-6) µm.
Secondary conidia were barrel shaped and hyaline with length (8- ) 9-12 (-12), width (5- ) 5-7 (-8).
Incase of resting spores Chlaymadospores were thick walled, dark brown 11-14 µm long (9-12) µm wide.
(Van Wyk, et al., 2007)
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All the isolates are morphologically compared which provides the cause is a fungi named asC. manginecans
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Sr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Results
GPS Values In vitro (Ceratocystis manginecans.)
Elevation North East1 26(a) 20-1-11 MZG, Qasba gujrat. 378' 30˚09.273" 70'57.355" 20-1-11 Yes2 27S(a) 20-1-11 MZG, Qasba gujrat 357' 30˚09.286" 70˚57.362" 20-1-11 Yes
Isolates
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Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporesW l Beak l Beak w Globosed
ascomaCylindrical Barrel
e.s w.s tip base w W l w l w l26a Muzzafergarh Yes 4 7 4 625 18 38 225 5 29 5 12 9 1227a Muzzafergarh Yes 4 7 3 650 22 28 237 5 20 6 10 10 13
Morphology
26a 27a
Perithecia Beak Tip
conidia
A.spore
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Sr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia
appearGPS Values (Ceratocystis manginecans.)
Flag No. Elevation
North East
3 28 25-3-2011 Talamba, Teh. Mian Channu
Distt. Khanewal.
324 424 30°31.097´
72°13.062´ 25-3-2011 5/4/2011 Yes
4 40 26-3-2011 Keekar wali Basti R.B. Thatha
Qureshi, Muzzafergarh
338 421 29°55.875´
71°10.497´ 26-3-2011 2/4/2011 Yes
Isolates
40
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28 Khanewal. Yes 4 7 3 555 25 35 200 5 23 7 12 9 11
40 Muzzafergarh Yes 4 7 4 575 17 35 183 5 29 5 10 9 12
Sample no.
Place c. manginecans Morphology
Ascospore Perithecia Conidia ChlaymadosporesW l Beak l Beak w Globosed
ascomaCylindrical Barrel
e.s w.s tip base w W l w l w l
Morphology
28 40
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5 46 26-3-2011 Habibabad, Muzzafergarh
345 327 30°04.550´
71°14.268´ 26-3-2011 2/4/2011 Yes
6 57 27-3-2011 Thana Alpaa, Multan.
358 383 30°19.728´71°31.207´ 27-3-2011 6/4/2011 Yes
Sr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia appearGPS Values (Ceratocystis
manginecans.)
Flag No. Elevation
North East
Isolates
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46 Muzzafergarh Yes 4 6 4 658 18 37 255 5 30 5 10 9 1157 Multan Yes 4 7 4 655 18 40 250 5 29 5 11 9 11
MorphologySample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporeW l Beak
lBeak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
46 57
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7 58 27-3-2011 Thana Alpaa, Multan.
359 380 30°19.724´ 71°31.227´ 27-3-2011 5/4/2011 Yes
8 59 27-3-2011 Thana Alpaa, Multan.
360 380 30°19.701´ 71°31.251´ 27-3-2011 4/4/2011 Yes
Sr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia appearGPS Values (Ceratocystis
manginecans.)
Flag No. Elevation
North East
Isolates
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58 Multan Yes 5 7 4 650 25 32 230 4 28 6.25 11.25 10 1359 Multan Yes 4.5 7 3 645 17 38 225 4 26 5 12 9 11
MorphologySample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporeW l Beak l Beak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
58 59
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9 61 27-3-2011 Baber Chock, Multan.
362 349 30°09.793´
71°32.346´ 27-3-2011 7/4/2011 Yes
10 63 27-3-2011 Baber Chock, Multan.
364 373 30°09.904´
71°32.302´ 27-3-2011 5/4/2011 Yes
Sr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia appearGPS Values (Ceratocystis
manginecans.)
Flag No. Elevation
North East
Isolates
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61 Multan Yes 4 7 3 625 22 25 188 5 28 5 10 9 12
63 Multan Yes 4.5 7.5 3 514 24 29 238 5.5 27 6 11 10 14
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporeW l Beak l Beak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
Morphology
6163
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11 64 27-3-2011 Trri aali bhaini, Multan.
365 336 30°09.718´
71°32.668´ 27-3-2011 5/4/2011 Yes
12 65 27-3-2011 Bhaini, Multan. 366 376 30°10.012´ 71°32.874´ 27-3-2011 5/4/2011 Yes
Sr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia appearGPS Values (Ceratocystis
manginecans.)
Flag No. Elevation
North East
Isolates
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64 Multan Yes 4 7 4 520 25 38 225 5 23 7 12 11 13
65 Multan Yes 5 7 3 263 26 33 238 6 30 5 10 9 11
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporeW l Beak l Beak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
Morphology
6465
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13 67 28-3-2011 11/8R-Makhdum pur, Teh. Kabeer
wala, Distt. Khanewal
369 412 30°27.610´72°05.816´ 28-3-2011 4/4/2011 Yes
14 68 28-3-2011 11/8R-Makhdum pur, Teh. Kabeer
wala, Distt. Khanewal
370 412 30°27.612´ 72°05.815´ 28-3-2011 6/4/2011 Yes
Sr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia appearGPS Values (Ceratocystis
manginecans.)
Flag No. Elevation
North East
Isolates
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67 Khanewal. Yes 4 7 3.5 650 23 30 230 5 25 5 10 10 12
68 Khanewal. Yes 5 7.5 3 660 24 40 225 5.5 29 5 10 10 13
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporeW l Beak l Beak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
Morphology
6768
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15 70 28-3-2011 11/8R-Makhdum pur, Teh. Kabeer
wala, Distt. Khanewal
372 425 30°27.595´ 72°05.807´ 28-3-2011 6/4/2011 Yes
16 71 28-3-2011 11/8R-Makhdum pur, Teh. Kabeer
wala, Distt. Khanewal
373 418 30°27.637´
72°05.791´
28-3-2011 4/4/2011 Yes
Sr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia appearGPS Values (Ceratocystis
manginecans.)Flag No. Elevatio
nNorth East
Isolates
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70 Khanewal. Yes 5 7 3 655 23 28 245 4.5 25 7 12 11 14
71 Khanewal. Yes 4.5 7 3 650 24 30 238 5 23 6 11 9 11
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporeW l Beak l Beak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
Morphology
70 71
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17 72 28-3-2011 11/8R-Makhdum pur, Teh. Kabeer
wala, Distt. Khanewal
374 420 30°27.647´ 72°05.776´ 28-3-2011 4/4/2011 Yes
18 73 28-3-2011 Satara Morr, Teh. & Distt. Khanewal.
375 398 30°30.455´ 72°11.050´ 28-3-2011 4/4/2011 Yes
Sr. No.
Sample No.
Date of sample
collection
Location of sample
collection
Date of isolation
Date of Perithecia appearGPS Values (Ceratocystis
manginecans.)
Flag No. Elevation
North East
Isolates
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72 Khanewal. Yes 4 7 3 625 17 35 225 5 22 7.5 12 9 11
73 Khanewal. Yes 5 7 4 650 15 38 240 5 25 7 11.5 9 12
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporeW l Beak l Beak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
Morphology
72 73
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19 74 28-3-2011 WaryamaAbad, Teh. &
Distt. Khanewal.
376 402 30°30.647´ 72°11.251´ 28-3-2011 5/4/2011 Yes
20 75 28-3-2011 Talamba road khanewal.
377 441 30°30.724´ 72°11.642´ 28-3-2011 4/4/2011 Yes
Sr. No.
Sample No.
Date of sample
collection
Location of sample
collection
Date of isolation
Date of Perithecia
appearGPS Values (Ceratocystis manginecans.)
Flag No.
Elevation North East
Isolates
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74 Khanewal. Yes 4.5 7.5 4 550 20 36 235 4 26 6 10 9 1475 Khanewal. Yes 4.5 7.5 4 525 25 25 252 5 33 6 11 10 13
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporeW l Beak l Beak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
Morphology
7475
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21 76 28-3-2011 Talamba road khanewal.
378 445 30°30.726´ 72°11.645´ 28-3-2011 7/4/2011 Yes
22 87 14-5-2011 Sher Shah (Toll plaza) Multan
D87 368 30°04.367´ 71°18.411´ 14-5-2011 28-5-2011 Yes
IsolatesSr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia
appearGPS Values (Ceratocystis manginecans.)
Flag No.
Elevation North East
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76 Khanewal. Yes 5 7 4 562 27 27 250 5 20 7 12 11 14
87 Multan Yes 4 7 3 538 25 34 250 4 22 6 10 10 12
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporeW l Beak l Beak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
Morphology
76 87
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23 88 14-5-2011 Sher Shah (Toll plaza) Multan
D88 363 30°04.371´ 71°18.425´ 14-5-2011 25-5-2011 Yes
24 91 14-5-2011 Sunaryanwali, Khan garh,
Muzzafergarh
D91 356 29°55.315´ 71°08.758´ 14-5-2011 29-5-2011 Yes
IsolatesSr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia
appearGPS Values (Ceratocystis manginecans.)
Flag No.
Elevation North East
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88 Multan Yes 5 7.5 4 525 25 33 225 5 29 6 11 12 14
91 Muzzafergarh Yes 5 7.5 4 600 25 38 200 5 30 5 11 11 13
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia Chlaymadospore
W l Beak l Beak w Globosed ascoma
w
Cylindrical Barrel
e.s w.s tip base W l w l w l
Morphology
8891
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25 92 14-5-2011 Sunaryanwali, Khan garh,
Muzzafergarh
D92 343 29°55.310´ 71°08.757´ 14-5-2011 28-5-2011 Yes
26 95 14-5-2011 Sunaryanwali, Khan Garh,
Muzzafergarh
D95 345 29°55.331´ 71°08.780´ 14-5-2011 3/6/2011 Yes
IsolatesSr. No.
Sample No.
Date of sample
collection
Location of sample
collection
Date of isolation
Date of Perithecia
appearGPS Values (Ceratocystis manginecans.)
Flag No.
Elevation North East
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92 Muzzafergarh Yes 4.5 7 3 514 23 35 250 4 20 6 10 11 14
95 Muzzafergarh Yes 4 7 3 575 17 33 253 5 20 5 10 9 12
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporesW l Beak l Beak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
Morphology
92 95
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27 104 16-5-2011 Union council bhao di
qureshan , RYK.
D104 272 28°36.847´ 70°16.312´ 16-5-2011 3/6/2011 Yes
28 105 16-5-2011 Union council bhao di
qureshan , RYK.
D105 275 28°36.832´ 70°16.313´
16-5-2011 6/6/2011 Yes
IsolatesSr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia
appearGPS Values (Ceratocystis manginecans.)
Flag No.
Elevation North East
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104 Rahim Yar Khan
Yes 5 7.5 3 525 20 25 250 5 23 7 10 10 13
105 Rahim Yar Khan
Yes 4 7 4 520 17 35 183 5 29 6 12 9 11
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporesW l Beak l Beak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
Morphology
104 105
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29 106 16-5-2011 Moza mohamadAli, RYK.
D106 283 28°39.529´ 70°15.721´ 16-5-2011 6/6/2011 Yes
30 108 16-5-2011 Kurr man singh, road moza shhna
wala, Dhaloon agri frm, RYK.
D108 335 28°36.373´ 70°13.618´ 16-5-2011 6/6/2011 Yes
IsolatesSr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia
appearGPS Values (Ceratocystis manginecans.)
Flag No.
Elevation North East
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106 Rahim Yar Khan
Yes 5 7.5 3 625 23 33 238 6 30 5 10 10 14
108 Rahim Yar Khan
Yes 5 7.5 4 550 20 25 200 5 24 5 12 9 14
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporesW l Beak l Beak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
Morphology
106 108
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31 109 14-9-2011 Mouza MohmadAli Magssi, RYK.
D109 295 28°37.963´ 70°15.897´
14-9-2011 27-9-2011 Yes
32 110 14-9-2011 Union council Bhao Di
Qureshan, RYK.
D110 293 28°36.716´ 70°17.739´
14-9-2011 29-9-2011 Yes
IsolatesSr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia
appearGPS Values (Ceratocystis manginecans.)
Flag No.
Elevation North East
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109 Rahim Yar Khan
Yes 4 7 3 520 20 25 225 5 22 6 11 10 13
110 Rahim Yar Khan
Yes 4 7 4 550 18 25 250 4 23 6 10 10 14
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporesW l Beak l Beak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
Morphology
109 110
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33 113 15-9-2011 Basti Bahram Baray Khoo Wali
Mouza Thul Ganga
Qureshan,, RYK.
D113 305 28°39.318´ 70°23.429´
15-9-2011 Yes
34 115 15-9-2011 Abad Pur, RYK. D115 288 28°39.212´ 70°22.542´
15-9-2011 29-9-2011 Yes
IsolatesSr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia
appearGPS Values (Ceratocystis manginecans.)
Flag No.
Elevation North East
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113 Rahim Yar Khan
Yes 4 7.5 3.5 600 20 28 225 5 18 5 10 9 12
115 Rahim Yar Khan
Yes 4 7 4 625 18 30 225 6 20 5 10 11 13
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporesW l Beak
lBeak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
Morphology
113115
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35 119 16-9-2011 Par Labbay Road , RYK.
D119 277 28°33.914´ 70°15.535´ 16-9-2011 Yes
36 121 17-9-2011 Bosan Farm KLP road, RYK.
D121 295 28°57.664´ 70°08.758´
17-9-2011 27-9-2011 Yes
IsolatesSr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia
appearGPS Values (Ceratocystis manginecans.)
Flag No.
Elevation North East
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119 Rahim Yar Khan
Yes 25 38 12 14
121 Rahim Yar Khan
Yes 5 8 4 675 22 38 250 6 25 6 12 10 12
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporesW l Beak l Beak w Globosed
ascomaw
Cylindrical
Barrel
e.s w.s tip base W l w l w l
Morphology
119121
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37 122 17-9-2011 JDW, Akhtarabad,
RYK.
D122 282 28°31.253´ 70°01.799´
17-9-2011 24-9-2011 Yes
38 125 17-9-2011 Kurr Maan Singh, RYK.
D125 305 28°36.146´ 70°13.675´
17-9-2011 24-9-2011 Yes
IsolatesSr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia
appearGPS Values (Ceratocystis manginecans.)
Flag No.
Elevation North East
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122 Rahim Yar Khan
Yes 4 7 3 675 20 45 275 5 30 7 12 9 13
125 Rahim Yar Khan
Yes 5 7.5 4 550 20 33 270 5 25 6 11 9 12
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporesW l Beak l Beak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
Morphology
122 125
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39 126 17-9-2011 Kurr Maan Singh, RYK.
D126 299 28°36.405´ 70°13.454´ 17-9-2011 24-9-2011 Yes
40 127 17-9-2011 Kurr Maan Singh, RYK.
D127 298 28°36.412´ 70°13.464´ 17-9-2011 24-9-2011 Yes
IsolatesSr. No.
Sample No.
Date of sample
collection
Location of sample collection
Date of isolation
Date of Perithecia
appearGPS Values (Ceratocystis manginecans.)
Flag No.
Elevation North East
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126 Rahim Yar Khan
Yes 4.5 7.5 4 600 15 45 188 4 20 7 11 10 13
127 Rahim Yar Khan
Yes 5 7.5 3 635 15 35 220 5 25 6 12 11 13
Sample no.
Place c. manginecans
Morphology
Ascospore Perithecia Conidia ChlaymadosporesW l Beak l Beak w Globosed
ascomaw
Cylindrical Barrel
e.s w.s tip base W l w l w l
Morphology
126 127
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In our study the range of morphological characters are as follows which is almost fall in the prescribed range of Van Wyk.
The ascomatal bases were globosed, black (153- ) 183-275 (-281) µm in diameter.
Ascomatal necks were dark brown to black becoming lighter at apices (514- ) 514-650 (-673) µm long, (25- ) 25-45 (-48) µm wide at base, (14- ) 15-26 (-26) µm wide at apices.
Ostiolar hyphae were hyaline and divergent (42- ) 45-62 (-69) µm long.
Ascospores were hyaline, hat-shaped (3-4) µm long, (4-5) µm wide excluding sheath and (7-8) µm wide including sheath.
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Primary cylindrical-shaped hyaline conidia were long (15- ) 18-32 (-33) µm wide (3- ) 4-6 (-6) µm.
Secondary conidia were barrel shaped and hyaline with length (8- ) 9-12 (-12), width (5- ) 5-7 (-8).
Incase of resting spores Chlaymadospores were thick walled, dark brown 11-14 µm long (9-12) µm wide.
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Further identification will be done on the bases of molecular characterization which is under progress.
Special thanks to Punjab Agriculture Research Board for funding this project.
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