David Doll

UC Agricultural and Natural Resources

UCCE Merced County
6-23-10

Dying Trees do Tell Tales:

Implications of Biological Agents
in Prunus Replant Disease.
David Doll
University of California Cooperative Extension
Merced County

G. Browne1, B. Holtz2, B. Lampinen3

CPGRU USDA-ARS, Department of Plant Pathology, UC Davis1,
UCCE Madera2, UC Davis Plant Sciences3

February12th, 2009
Southern San Joaquin Valley Almond Day
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

Replant Disease of Almond

Symptoms on
almond:

Healthy (L) and

PRD-affected almond trees
(R) in Butte County, 2003

CP-fumigated plot Non-fumigated plot

 David Doll

Responses to spot, strip, and

UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

broadcast fumigation treatments

(Location: Firebaugh, pre-plant fumigated Oct 2006, planted Jan 2007)
Fum.
per Disease Trunk
Treated area in tree orch. severity diameter
Tr Fumigant, rate per treated row acre rating February
t. areaa (and % of total area) (lbs) 6/20/07 2008 (mm)
1 Control None 0 1.8 20
2 Methyl bromide, 400 lb/a 8-ft strip (38%) 152 0.8 24
3 Telone II, 350 lb/a 8-ft strip (38%) 133 1.0 27
4 Chloropicrin (CP), 400 lb/a 8-ft strip (38%) 152 0.1 38
5 CP, 300 lb/a 8-ft strip (38%) 114 0.4 37
6 CP, 200 lb/a 8-ft strip (38%) 76 0.1 39
7 CP, 400 lb/a 8x8-ft tree sites (17%) 68 0.5 34
Midas (IM:CP. 50:50), 300
8 8-ft row strip (38%) 152 0.3 36
lb/a
9 Telone C35, 550 lb/ac 8-ft row strip (38%) 209 0.1 36
10 Pic-clor 60, 550 lb/ac 8-ft row strip (38%) 209 0.0 39
11 Pic-clor 60, 400 lb/ac 8-ft row strip (38%) 152 0.3 35
12 Telone C35, 550 lb/ac 8x8-ft tree sites (17%) 93 0.3 33
13 Telone C35, 550 lb/ac Broadcast (100%) 550 0.1 37
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

Replant Disease of Almond

Healthy RD-affected

Healthy (L) and replant disease-affected (R) almond trees,

Madera County 2007
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

Effects of replant disease on root system

Root length densities at 0 to 2 ft depth; determined by root

excavation and digital imaging
Lovell peach rootstock Marianna 2624 rootstock
Total root length per sample (cm)

4000 4000

3000 Control 3000 Control

Pre-plant chloropcirin
Pre-plant chloropcirin
2000
2000

1000
1000

0
0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 5
0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 Root diameter (mm)
Root diameter (mm)
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Probesting, E.L., and Gilmore, A.E. 1941. The relation of peach root toxicity to
re-establishing of peach orchards. Proc. Am. Soc. Hort. Sci. 38:21-26.
Probesting, E.L. 1949. The peach replant problem. Hort. News 30:2213-2214.
Probesting, E.L. 1950. A case history of a "peach replant" situation. Proc. Amer. Soc.
Hort. Sci. 56:46-48.
Patrick, Z. A. (1955). "The peach replant problem in Ontario. II. Toxic substances
from the microbial decomposition products of peach root residues."
Canadian Journal of Botany 33: 461-486.
Wensley, R. N. (1956). "The peach replant problem in Ontario. IV. Fungi
associated with replant failure and their importance in fumigated and
nonfumigated soils." Canadian Journal of Botany 34: 967-982.
Mountain, W. B. and H. R. Boyce (1958). "The peach replant problem in Ontario.
V. The relationship of parasitic nematodes to regional differences in
severity of peach replant failure." Canadian Journal of Botany 36: 125-
134.
Gilmore, A.E. 1959. Growth of replanted peach trees. Proc. Am. Soc. Hort. Sci.
73:99-111.
Hine, R.B. 1961. The role of amygdalin breakdown in the peach replant problem.
Phytopathology 51:10-13.
Hine, R.B. 1961. The role of fungi in the peach replant problem. Plant Disease
Reporter 45:462-466.
Gur, A., Cohen,Y. 1989. The peach replant problem- some causal agents. Soil Biol.
Biochem. 21:829-834.
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Peach Root Toxicity:
-Allelopathy

Nematodes:
-Ring, Rootknot, etc.
Microbial involvement:
-Bacteria
-Fungi
No such thing:
Just a growth response to
fumigation
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Control MBr 400 lb/A MBr 2700 lb/A

• Soil from old peach

orchard

• Pre-plant fumigation
treatments applied to
microplots before
planting with Nemaguard

Chloropicrin 400 lb/A Chloropicrin 2700 lb/A

At end of first growing season, Nemaguard peach seedlings

 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Peach Root Toxicity:
-Allelopathy

Nematodes:
-Ring, Rootknot, etc.
Microbial involvement:
-Bacteria
-Fungi
No such thing:
Just a growth response to
fumigation
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Nemaguard Seedlings planted in soils that have been previously cropped in
peaches or grapes for 15 years. Microplot trials.

90
Min. sig. dif. = 24 Min. sig. dif. = 15
80
Maximum Shoot Length (cm)

70
60
Control
50
Methyl Bromide
40 Chloropicrin
30 Telone II
Telone C35
20
10
0
Peach Soil Grape Soil
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Nematodes:
-Ring, Rootknot, etc.
Microbial involvement:
-Bacteria
-Fungi
No such thing:
Just a growth response to
fumigation
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Nematode Involvement in PRD:
• Butte County orchard RD trials (2001-2004):
– No significant counts of plant parasitic nematodes (3 orchards)
– Sugar floatation and mist chamber extractions

• Parlier microplot trials (2002-2005):

– Only pin nematode (Paratylenchus sp.) present in significant number
(3 repeated trials); not correlated with RD incidence

• Madera County orchard trials (2003- ):

– To date, minor or no nematode involvement; sampling continuing

• Conclusion:
– RD and nematode parasitism not the same, they are distinct replant
problemswith biological agent(s) other than plant parasitic nematodes
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Nematodes:
-Ring, Rootknot, etc.
Microbial involvement:
-Bacteria
-Fungi
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Hypotheses, Prunus replant disease (PRD):
 PRD is induced by unfavorable shifts in soilborne microbial
communities associated with previous cultivation of the
host
 The shifts can be detected by complementary culture-based
and culture-independent sampling of fine roots and the
rhizosphere
 Community members contributing to (or suppressing)
incidence of PRD can be cultured and tested for
pathogenicity (or disease suppression)
 Effects of semi-selective soil treatments on incidence and
severity of PRD will provide additional evidence for
microbial roles in the disease
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Roots from healthy tree Roots from RD-affected tree

Roots and soil from healthy and RD-affected trees

Whole-organism assays Molecular assays Soil treatments

• Direct quantification (nematodes) • DNA fingerprinting (PCR of • Semi-selective
• Isolation and quantification in culture rDNA; fungi, bacteria, chemicals or
(fungi, bacteria) • cloning of fragments, DNA- treatments in
sequence-based id) Microplot, GH
tests
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Effects of semi-selective soil treatments
Greenhouse trial:

No heat Autoclaved
50
Discolored roots (%)

40

30

20

10

0
il
ol

.
d

r
am

u
om
en
tr

ac
ax
on

or
id

id

em
M
iv

hl
C

R
D

N
C
pt.+
re
St
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County

What causes replant disease?

6-23-10

Effects of semi-selective soil treatments

Microplot trial:
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Effects of semi-selective treatments on severity of PRD,
microplots
d

a
bc
d

cd

bc
ab

Top fresh wt. of plants per plot (g)

 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Hypotheses, Prunus replant disease (PRD):
 PRD is induced by unfavorable shifts in soilborne microbial
communities associated with previous cultivation of the
host
 The shifts can be detected by complementary culture-based
and culture-independent sampling of fine roots and the
rhizosphere
 Community members contributing to (or suppressing)
incidence of PRD can be cultured and tested for
pathogenicity (or disease suppression)
 Effects of semi-selective soil treatments on incidence and
severity of PRD will provide additional evidence for
microbial roles in the disease
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Roots from healthy tree Roots from RD-affected tree

Roots and soil from healthy and RD-affected trees

Whole-organism assays Molecular assays Soil treatments

• Direct quantification (nematodes) • DNA fingerprinting (PCR of • Semi-selective
• Isolation and quantification in culture rDNA; fungi, bacteria, chemicals or
(fungi, bacteria) • cloning of fragments, DNA- treatments in
sequence-based id) Microplot, GH
tests
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County

What causes replant disease?

6-23-10

Ordination of bacterial incidence, Parlier trial, 2004

1.0
Control*Bleach

Methyl Bromide*Bleach
Rhiz-1
RhizA-1 Control

RhizA-2
Pseud-2 Pseud-1

Entero Methyl Bromide

Flav-4
Chloropicrin*Bleach Microb-1
Bacill-6
Vario-1 Control*Rinse
Microb-3
Chloropicrin Pseud-3 Arthro-2
Bacill-4
Methyl Bromide*Rinse
• N=266 Chloropicrin*Rinse

• Ordination significant
at P=0.008 for axis 1
2003 Trial Year
-1.0

and P=0.002 for axis 2.

-1.0 1.0
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County

What causes replant disease?

6-23-10

Ordination of cultured fungal incidence, Firebaugh replant trial, 2007

1.0
Control*Rinse

Fusoxy1
Asper
Fus3 Fus4
Unk-C Acremon

CP*Bleach
Control CP Tricho-2

CP*Rinse

Cylind
• N=150
Fus1
• Ordination
significant at
-1.0

Control*Bleach
P=0.002 for each
axis -1.0 1.0
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County

What causes replant disease?

6-23-10

Testing pathogenicity of bacteria and fungi on

Nemaguard peach
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Pathogenicity of bacteria associated with replant disease on NG peach
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Pathogenicity of fungi associated with replant disease on NG peach
Disease serverity score
(0=healthy, 5=dead)
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

What causes replant disease?

Microbial involvement:
-Bacteria
-Fungi
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County

Non-Fumigant Management of 6-23-10

Replant Disease
Application of pre-plant cropping and fallow treatments.
Microplot experiments.
 David Doll
UC Agricultural and Natural Resources

Non-Fumigant Management of
UCCE Merced County
6-23-10

Replant Disease
Responses of Nemaguard peach seedlings to short-term
fallow, rotation treatments. Microplot Experiments.
Top fresh wt. end of first 800

600
year (grams)

400

200

0
 David Doll
UC Agricultural and Natural Resources

Non-Fumigant Management of
UCCE Merced County
6-23-10

Replant Disease
The affects of Piper-Sudan on replant disease of peach. Field
Trials. Planting: O’Henry Peach on Nemaguard following 20
yr old Santa Rosa Plums on Marianna 2624
 David Doll
UC Agricultural and Natural Resources

Non-Fumigant Management of
UCCE Merced County
6-23-10

Replant Disease
Responses to spot, strip, and broadcast fumigation treatments, Firebaugh
(pre-plant fumigated Oct 2006, planted Jan 2007)
Increase
Fumigant Fumigant in trunk
per per Disease severity diameter
treated orchard rating by 30 Aug
Fumigation treatment acre acre Sudan grass rotation (0 to 5 scale) 2008
(Oct 2007) (lbs) (lbs) (Jul-Sep 2007) (7 Jul 2008) (mm)
no 1.5 7.9
Control 0 --
yes 0.9 14.1
MB, by conventional no 0.9 22.6
shanks to 8’-wide row 400 168
strips yes 0.5 20.4
Telone C35, by no 0.3 28.3
conventional shanks to 8’- 540 227
wide row strips yes 0.3 31.9
Telone C35, by GPS- no 0.7 21.4
controlled shanks to 5x 6’ 540 81
tree spots yes 0.5 24.2
Inline, by single drip no 0.8 20.8
emitters 540 43
to 4’-dia. tree spots yes 0.6 21.3
Chloropicrin, by GPS- no 0.6 24.2
controlled shanks to 5x6’ 400 60
tree spots yes 0.2 26.5
None, yeast extract root no 1.2 10.4
spray and drench at 0 --
planting yes 1.0 14.0
Minimum significant difference (according to 95% confidence intervals): 8.9
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County

Non-Fumigant Management of 6-23-10

Replant Disease
22 a
Tree Diamter (mm) 21.5
21
20.5
20
19.5
b
19
18.5
18

Control Sudan
P=0.048
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

Conclusions:
 Replant disease (RD) of almond is a biologically induced
disease showing poor growth or survival in almond
planted after Prunus.
◦ Prevented by pre-plant fumigation with fumigants containing
Chloropicrin.

 Some fungi and bacteria are being implicated, multiple

approaches are being used to determine causal agents
and developing greenhouse assays.

 An unfavorable microbial community may be

responsible for the disease:
◦ Shifting the microbial community in favor of the newly planted
Prunus sp. tree may provide some level of disease control.
◦ Pre-plant application of treatments or cover cropping may
provide alternative control reducing the use of fumigants
 David Doll
UC Agricultural and Natural Resources
UCCE Merced County
6-23-10

Thank you!
Almond Board of California

USDA-ARS Pest Mgt. Program for Integrated

Alternatives to Methyl Bromide

TriCal, Inc.

Commercial almond and peach growers of CA

Multiple scientific personnel involved