American hazelnut micropropagation
Transcript of American hazelnut micropropagation
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Eric Zeldin,
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(Fig. 1).
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own at the Ho
ersity of Wisc
king with this
ver thirty yea
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ROPROPAG
R THE AME
Scientist, Uni
August
challenges in
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program of P
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elnut.
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GATION PO
ERICAN HA
iversity of Wi
t 14th, 2012
n the develop
als. In most fr
is not feasible
s that will byp
the American
th a low perce
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plication and
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s been used t
any recalcitra
Professor Bre
epartment of
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dy plant biote
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nt Initiative, w
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s and disease
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ow and estab
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onnel require
pagation rese
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ting the Amer
OTENTIAL
AZELNUT
sconsin – Ma
ment of the A
ruit and nut c
e with the Am
pass the desir
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uccessful and
d. Research i
arch with the
search (mulch
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h replicated c
rican hazelnu
adison
American haz
crops, selecte
merican hazel
red individual
Fig. 1. One ro
10% satisfacto
s and very few
mate and loc
ncrease once
an those obt
d it can be qu
is necessary t
e American ha
hing, fertilizin
e the selectio
clonal trials. T
ut, from select
zelnut as a ne
ed individuals
nut because g
l anyway. Ro
ooting test wit
ory rooting.
w drawbacks
cation.
cultures are e
ained from co
ite expensive
to overcome
azelnut at thi
ng, pruning, e
on and develo
This article de
ted parent pl
ew crop is the
are grafted t
grafting is dif
oted cuttings
th American ha
to microprop
established.
onventional c
e due to the c
these difficul
is point in tim
etc.) uncompr
opment of eli
escribes in tex
ants to hund
e difficulty in c
to produce ne
fficult and the
s can be utiliz
azelnut that yi
pagation. Wit
cuttings.
costs of specia
lties.
me are: 1) to p
romised by th
te individuals
xt and picture
reds of genet
clonally
ew plants tha
e seedling
ed in many
ielded only
th
alized
perform
he genetic
s for
es our initial
tically
t
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regar
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opropagation
s to handle th
oorganisms u
no contamin
esh medium (
d multiplicatio
e considered
mization is im
e enough mic
entional cutt
wing plantlets
er set of cuttin
h tree bands
e plants had
ficant as we w
to produce an
ortant for the
of the lower
ds and heavy f
ts are very vig
eld‐establish
plants have st
ed above. We
be begin to ex
growth after
have started t
ral selections
ormance trial
opropagation
American haz
west. Eventu
opropagated
nsive and res
ned.
pictures below
ave obtained
rding micropr
ed with the se
hern Wiscons
lected plants
ound collar re
n requires ste
e tissue (Fig.
using dilute bl
ates) develop
(Fig. 4). The b
on (Fig. 5). So
it more impo
mportant for t
rocuttings we
ings (Fig. 6).
was readily a
ngs were root
and after fur
excellent roo
wanted to ma
n undergroun
later produc
stems were b
fertilization r
gorous and ar
ment trials us
tarted to bran
e have produc
xamine the ef
planting.
to work on th
s in vitro that
s in 2013 or 2
n will play a vi
elnut for a ne
ally other kin
plants as sto
search aimed
w offer a clea
d so far. If the
ropagation of
election of hi
sin and Micha
and establish
egion (Fig. 2).
rilized culture
3). In additio
each or simil
ped new grow
bases were al
o far the proc
ortant to test
he improvem
ere available,
The quality o
acclimated to
ted, acclimate
rther growth w
ot mass and o
aintain viable
nd vegetative
tion of new s
buried when t
esulted in ext
re quickly app
sing fall plant
nch and it is e
ced a sufficie
ffects of diffe
he second stat
should be rea
2014. We are
ital role in the
ew orchard cr
ds of propaga
ck plants may
at such seco
ar view of the
ere are any co
f the America
gh nut produ
ael Demchik i
hed in the gre
Because bac
e vessels and
on, the startin
ar chemicals
wth from late
lso transferre
cess has work
rooting, plan
ment of micro
rooting was
of roots produ
o ambient con
ed and estab
were acclima
ver time the
vegetative b
axis for the e
tems which m
the plants we
tremely unifo
proaching a s
tings of dorm
encouraging t
nt number of
erent levels of
ted goal: we
ady for replic
e certain that
e future impr
rop for the Up
ation using
y prove to be
ndary propag
e process and
omments or q
an hazelnut, p
cing plants fr
n Central Wis
eenhouse. Th
cteria and fun
medium, as w
ng plant mate
to surface ste
ral buds and
ed, yielding ne
ked well, but w
nt developme
propagation e
tested in the
uced was exce
nditions by gr
lished in the g
ted to outsid
buds on the l
uds that wou
establishmen
may be critica
ere re‐potted
orm, rapidly g
ize suitable fo
ant plants an
to see branch
f plants to als
f mulch and d
now have
cated
rovement of
pper
less
gation is
the results
questions
please contac
rom the wild,
sconsin. Colla
hese were cu
ngi grow so fa
well as a ster
erial must be
erilize the tiss
when tall eno
ew stems; rep
we have not y
ent and field p
efficiency and
lab and foun
ellent and ne
adual exposu
greenhouse.
de conditions,
lower stems b
uld be buried
t of a collar‐l
al for nut prod
into 4 x 4 x 1
growing plant
or field planti
nd spring plan
hing from bur
so start to add
different leve
ct Eric Zeldin b
performed b
ar (basal, und
t back to forc
ast and are un
rile transfer c
isolated from
sue. Successf
ough, the tips
petition of th
yet optimized
performance
d thus final p
nd to be far m
ew growth on
ure to open a
These were
, initially unde
began to swe
with subsequ
ike region sim
ductivity ove
10 inch tree b
ts (Figs. 10 an
ing (Fig. 12).
ntings of activ
ried buds (Fig
dress goal 1 l
ls of fertilizer
by email at el
by cooperator
derground ste
ce new growt
ndesirable an
abinet and st
m contaminat
ful isolates (s
s were cut an
his process res
d the multipli
first. Howev
er plant cost.
more successf
the resulting
ir (Fig. 7). Su
potted into 2
er shade clot
ell (Fig. 9). Th
uent re‐pottin
milar to seedl
r time. There
bands. The lar
nd 11). Curren
These plants
vely growing p
. 13) for the r
isted above: i
r on plant est
lzeldin@wisc
rs Jason
em) divisions
th from the
nyway,
terilizable
ting
stem pieces
nd transferred
sulted in
cation phase
er,
.
ul to that of
g rapidly‐
bsequently, a
2 5/8 x 2 5/8 x
h (Fig. 8).
is was
ng. The idea
ings. This is
efore, a large
rger tree
ntly the
s will be used
plants.
reasons
in 2013 we
ablishment
.edu.
d
a
x
Fig. 2. Micropropagation starts with a high nut producing selection from the wild established in the greenhouse. Pruning of the
stems removes apical dominance and new, more juvenile stems erupt from the underground collar region (right picture).
Greenhouse stems are cleaner than those from the field (fewer bacteria and fungal spores) and juvenile tissue usually yields the
bests results for micropropagation.
Fig. 3. A sterile transfer cabinet used to perform sterile transfers. Stems from hazelnuts cultures (left in picture) will be cut and
placed on new media (center) using surgical steel quality sterilized tools (right).
Fig. 4. A hazelnut isolate exhibiting new growth (left picture) and a group of new stems produced in vitro (right picture).
Fig. 5. Hazelnut cultures in the culture room. The first three trays (left picture) contain about 500 stems genetically identical to
the source plant they were originally isolated from. A closer view (right picture) shows a culture with microstems ready for
cutting and rooting. Further optimization of the multiplication phase will result in faster and cheaper multiplication.
Fig. 6. Microcutting rooting trial. Two to three inch microcuttings were dipped in rooting hormone and placed in propagation mix
(peat/perlite) in a 1020 tray covered with a moisture retaining plastic dome. Greater than 95% rooting has been achieved using
microcuttings and rooting hormone treatments.
Fig. 7. Rooted microcutting with roots exposed (left picture) and young, greenhouse acclimated plants derived from microcuttings
(right picture). Good quality rooting is essential for the tissue to acclimate from protected, in vitro culture to exposed ambient
conditions.
Fig. 8. Greenhouse acclimated plantlets were repotted into 2 5/8 x 2 5/8 x 5 inch tree bands using commercial grower's mix and
subsequently acclimated to outside conditions in a coldframe.
Fig. 9. A plant from Fig. 8 with the tree band removed to show root development (left picture) and a detail of the lower stem of
another plant to show lateral bud swelling (right picture). These buds will be buried after re‐potting.
Fig. 10. Re‐potting of micropropagated plants to larger, 4 x 4 x 10 inch tree bands (left picture) and a top view of the re‐potted
plants showing a high degree of uniformity (right picture).
Fig. 11. A side view of the micropopagated plants in large tree bands after further growth.
Fig. 1
mont
(dorm
Fig. 1
comin
micro
new s
12. A single pla
th or so before
mant) and spri
13. Many of th
ng from buried
opropagated A
stems and nut
ant from the p
e dormancy se
ng 2013 (activ
he micropropag
d nodes (right
American haze
t producing cap
picture in Fig. 1
ts in. These m
ve growth) plan
gated plants a
picture). The
lnuts, as the re
pability.
11, demonstrat
micropropagate
nting to determ
are beginning t
development
e‐establishme
tes the strong
ed American h
mine which pla
to branch at th
of new stems
nt of a collar‐l
g, vigorous grow
hazelnut plants
anting time yie
his time (left p
s from buried n
like region ma
wth achieved
s will be tested
elds the best f
picture) and ma
nodes may be
y be required
to date, with a
d both for fall
field establishm
any of these b
a critical facto
for continual p
another
2012
ment.
branches are
or for
production of