FIELD PERFORMANCE OF BLUE PINE (PINUS WALLICHIANA) SEEDLINGS INOCULATED WITH SELECTED SPECIES OF BIO-INOCULANTS UNDER NURSERY CONDITIONS

1. FIELD PERFORMANCE OF BLUE PINE (PINUS WALLICHIANA)
SEEDLINGS INOCULATED WITH SELECTED SPECIES OF BIO-INOCULANTS
UNDER NURSERY CONDITIONS (Asif et al.,)
SUBMITTED BY : UDAY KUMAR LODH
MSc Forestry
ICAR JRF (AIR-2)
SUMITTED TO : Dr. GURUDATT HEGDE
FBT DEPARTMENT

2. INTRODUCTION
• Pinus wallichiana is commonly known as kail, blue
pine or Bhutan pine, is an evergreen large conifer tree
which has bluish feathery foliage.
• In the Himalayan region, kail is frequently found
between 1500-3000m.
• From Afghanistan in the west, the kail region
extends upto Bhutan and Arunachal Pradesh in the
east, although it is absent in considerable portions of
Kumaon and Sikkim. Other important places where
kail grows abundantly in the sub-continent are from
Garhwal through Jaunsar, the Shimla hills, Kulu,
Chamba and Muree hills.
• It is also a pioneer and strong light demanding
species.
• It can resist moderate shade for sometime but makes
little progress and ultimately fails to recover.It can
resist the wind by its massive root system.
• To meet the challenges like poor regeneration,
deforestation and spread of wastelands, introduction
of microbial inoculants at the nursery stage of forest
trees has become inevitable

3. OBJECTIVE
• To study Various growth characters viz., shoot height, collar diameter,
root length, and seedling survival at various intervals responded
significantly to all the microbial inoculants.
Pisolithus tinctorius

4. Laccaria laccata Azotobacter

5. Azospirillum
Pseudomonas fluorescens

6. MATERIALS AND METHODS
• The present investigations were undertaken at the Forest Nursery of Department of Forestry, Faculty of Agriculture and
Regional Research Station, SKUAST-Kashmir, Wadura, Sopore during 2009 and 2010.
• Microbial inoculants isolated from rhizosphere of blue pine forest stands were used in the studies.
Mass production of microbial inoculants: The two free living aerobic nitrogen fixing bacteria viz., Azotobacter sp and
Azospirillum sp were mass cultured using nutrient medium enriched with glucose and peptone. Plant growth promoting
rhizobacteria (PGPR)viz., Pseudomonas fluorescens and Bacillus subtilis were mass propagated in King’s B nutrient
broth. The two ectomycorrhizae viz., Pisolithus tinctorius and Laccaria laccata were mass multiplied in Melin Norkran’s
nutrient broth and Potato Dextrose Agar, respectively.
Field operations: For the microbial inoculation, one year old seedlings of blue pine of uniform heights and collar diameter
growing in polyethylene bags (9″ x 7″) containing 1 kg potting material of soil and sand mixture in the ratio of 1:1 were
selected.
Microbial inoculation: For inoculation, the different broth cultures of N-fixers, P-solubilizers and ectomycorrhizal
inoculants isolated from local forest stands were applied to the potting material (25 ml/seedling) in the month of March,
2009, without disturbing the root system of the seedlings.
Nursery operations: The seedlings were irrigated with rose-cans as and when needed and maintained virtually weed free by
manual weeding.
Plant growth measurement: All the growth parameters viz., plant height (cm), collar diameter (mm), seedling survival (%)
and root length (cm) were measured at an interval of 2 months up to 12 months. All the growth parameters of the seedlings at
the initial stage of the experiment were recorded

7. Table-1
Impact of microbial inoculation on plant height (cm) of Blue pine
(Pinus wallichiana ) at nursery stage
Results:
Azotobacter and Azospirillum inoculation exhibited 37.17 and 36.83percent more plant height over control.
Similarly Pseudomonas flourescens and Bacillus subtilis inoculation resulted in 35.56 and 33.91percent more plant height
while as the inoculation with two ectomycorrhizal fungi viz., Pisolithus tinctorius and Laccaria laccata resulted in 42.97
and 40.77 per cent more plant height as compared to control respectively
Moreover, there was an increasing trend in plant height from April to October and from December onwards till
February there was a slight increase.
(Asif et al.,)

8. DISCUSSION & JUSTIFICATION
• The increase in shoot height by P. tinctorius and L. laccata could be attributed to the
production of growth promoting substances like auxins (Dehn, 1982) and enhancement of
water absorption and nutrient mobilization (Dar et al., 1997) by vastly increased surface
area network of the fungal mycelia (Myer, 1992).

9. CONTROL
BS
PS
AS
AZ
LL
PT
PT = Pisolithus tinctorius;LL = Laccaria laccata;AZ = Azotobacter sp.; AS = Azospirillum sp.; PS =
Pseudomonas fluorescens; BS = Bacillus subtilis

10. Table-2
Impact of microbial inoculation on collar diameter (mm) of Blue pine
(Pinus wallichiana ) at nursery stage.
. The inoculation with P. tinctorius showed 16.87percent increment over control.
This was followed by Laccaria laccata (15.84%), Azotobacter (13.70%), Azospirillum (12.82%), Pseudomonas
fluorescens (9.81%) and Bacillus subtilis (8.84%), respectively.
The application of ectomycorrhizal fungi viz. P. tinctorius resulted in maximum collar diameter which was 16.87 per
cent more than control.
Moreover, plant collar diameter revealed a significant increase from April to October and thereafter till February
there was a slight non- significant increase
(Asif et al.,)

11. • . Enhancement in collar diameter could be due to the release of plant
growth substances and increase in the nutrient availability in the
root zone.
DISCUSSION & JUSTIFICATION

12. Table-3
Impact of microbial inoculation on seedling survival (%) of Blue pine
(Pinus wallichiana) at nursery stage.
The best results with respect to seedling survival percentage were obtained with the inoculation of
Pisolithus tinctorius which was 21.06 per cent more as compared to control
(Asif et al.,)

13. DISCUSSION & JUSTIFICATION
• Enhancement in seedling survival could be attributed to the ability of
microbial inoculants to secrete antifungal antibiotics, uptake of nutrients
by converting them into available forms and greater access to water and
production of growth promoting substances.
• The decrease in survival percentage of the species in winter months
could be attributed to low and below freezing soil temperatures which
might have stopped the growth of inoculants and other soil microflora
present there.

14. Tab1e-4
Impact of microbial inoculation on root length (cm) of Blue pine
(Pinus wallichiana) at nursery stage
Pseudomonas fluorescens, Bacillus subtilis and L. laccata inoculations resulted in 15.88, 14.24 and 18.21 per cent more
root length over control.
The application of Pisolithus tinctorius resulted in maximum root length which was 19.06 per cent more than control
and thus proved superior over all other microbial inoculants
(Asif et al.,)

15. DISCUSSION & JUSTIFICATION
• The increase in root length due to microbial inoculants could be
attributed to their capability to synthesize biologically active
substances and increased uptake of essential macronutrients.

16. • A pot experiment was carried out during 2009 and 2010 to study the field performance of blue
pine seedlings inoculated with selected species of bio-inoculants under nursery conditions .
• The experiment was laid in Completely Randomized Design with three replications which
comprised of seven inoculants (Azotobacter sp., Azospirillum sp., Pseudomonas fluorescens,
Bacillus subtilis, Pisolithus tinctorius, Laccaria laccata and control).
• Various growth characters viz., shoot height, collar diameter, root length, and seedling survival
at various intervals responded significantly to all the microbial inoculants.
• Among microbial inoculants the two mycorrhizae viz., Pisolithus tinctorius and Laccaria
laccata proved beneficial for all growth parameters than rest of the inoculants. It was
followed by Azotobacter sp., Azospirillum sp., Pseudomonas fluorescens and Bacillus subtilis.
• However for root length Pseudomonas fluorescens and Bacillus subtilis gave best results
than Azotobacter sp. and Azospirillum sp.
• Microbial inoculation of Pisolithus tinctorius and Laccaria laccata gave best results with respect
to per cent decrease in seedling mortality rate of the species.
• Thus the two treatments viz; Pisolithus tinctorius and Laccaria laccata proved superior for all
the studied growth parameters.
• Their findings show that the application of bio-inoculants improve the growth attributes of
blue pine seedlings under natural field conditions.