1. Mature leaves of the sponge gourd (Luffa cylindrica) can be used for transient gene expression through agroinfiltration. The leaves easily absorbed Agrobacterium tumefaciens containing a GUS reporter gene and quickly diffused it through the leaf tissue. 2. Compared to other cucurbit leaves, Luffa cylindrica leaves have large air spaces in the mesophyll that allow for better absorption and diffusion. Stomata structure is similar across cucurbit leaves but Luffa has larger sub-stomatal air spaces. 3. Following agroinfiltration, GUS expression was detected in Luffa leaf extracts and ph

1. Smooth Gourd Vegetable
Sponge, Wash Sponge,
Gourd Towel, Dishcloth
Gourd, Loofah Gourd
SPONGE GOURD
Submitted by
S.ADHIYAMAAN (2017603401)
I-M.Sc. VEGETABLE SCIENCE
DEPT. OF VEGETABLE CROPS
HC & RI, TNAU, CBE.- 641 003

2.  Botanicalname : Luffa cylindrica
 Family : Cucurbitaceae
 Chromosome number : 2n=26
 Origin : Sub-tropical Asian (India)

3. Area and production
 Sponge gourds are grown as mixed cropping in the river beds
and as mono crop in the garden lands, hence the exact area and
production are unknown.
 Nevertheless the estimated area under all the gourds is 4.05
lakh hectares in our country.

4. Economic importance
 Cultivated both on a commercial scale and in kitchen gardens
throughout India.
 as spring-summer and rainy season crops.
 The genus derives its name from the product 'loofah', which is used
in bathing sponges, scrubber pads, doormats, pillows, and
mattresses and also for cleaning utensils.
 Both the species contain a gelatinous compound called luffein.
 sponge gourds have some medicinal uses.

5.  Fruits of ridge gourd are ribbed and is called kali tori
whereas fruits of sponge gourd are smooth and is
called ghja tori.
 The sponge gourd fruits contain higher protein and
carotene than ridge gourd.
 The fiber of dry fruits is also used in a number of
ways.

6.  In South, southeast, and East Asia, the tender fruits
are eaten fresh or more commonly cooked and
consumed as a vegetable.
 Sometimes, the tender leaves and growing shoots are
also used as a pot herb.

7. SOIL
 The crops can be grown on a variety of soil types.
 Loamy soils are considered best if there is a good amount of
organic matter and if it is well drained.
 Soil should have good moisture-holding capacity especially in
summer season.
 A well drained soil having pH 6.0-7.5, good fertility and high
organic matter are ideal for cultivation of ridge and sponge
gourds.

8. Climate
 The crops are well adapted to a fairly wide range of
climatic conditions.
 Luffa requires a long warm season for best
production.
 It also grows best during the rainy season.
 Optimum temperature requirement for these crops is
25°-27°C.
 Due to its hard seed coat, there is a problem with
seed germination when the temperature is low.
 Excessive rainfall during the flowering and fruiting
period reduces the yield considerably.

9. Pusa Chikni
 A selection from Bihar collection.
 Early fruiting variety, flowering in about 45 days.
 The fruits are smooth and dark-green colour, more or less
cylindrical, 15-20 fruits per vine, suitable for both spring-
summer and rainy seasons.
 Released by Indian Agricultural Research Institute, New Delhi.
Pusa Sneha:
 Suitable for long distance transport, fruits are dark green, 20-
25 cm long with hard skin and soft flesh.
 Yield 12 t/ha.

10. Phule Prajakta:
 It is a selection from local germplasm. It is released
by Mahatma Phule Krishi Vidyapeeth, Rahuri
(Maharashtra).
 It is suitable for summer and kharif season. Fruits are
medium green, flesh white, dark-green.
 Yield -15tonnes/ha.

11. Pusa Supriya:
 It is released by Indian Agricultural Research Institute, New
Delhi.
 It is suitable for summer and kharif seasons.
 Fruits are distinct, pale-green, medium- long, 15-20cm long,
straight and slightly curved at the stem end, pointed distal- end
with long peduncles, smooth, non-hairy, slim, narrow green
linings and spots, tender endocarp, skin very thin, on an
average 10-12 fruits weighing a kg.
 The fruits are ready for first picking at 47-55 days after sowing.
 A fruit weigh about 110g.
 It is an improvement over Pusa Chikni.
 It gives a 220% more yield than Pusa Chikni.
 It yields 8-9 tonnes/ha during spring-summer and 10-11
tonnes/ha during kharif season.

12. Season
 The time of planting usually depends upon the
season, location and market demand.
 The crop is grown in both the summer and rainy
seasons.
 Summer season crop is sown in January-February and
rainy season crop is sown in June-July.
 In areas having mild winters sowing can be done in
October, which gives an early crop in March.
 In river beds, in northern India, sowing is done in
November- January.

13.  Cropping systems: In garden land areas there is a definite
systems of growing cucurbits in rotation and mixed cropping.
 Preparation of land
 The land should be prepared to obtain the tilth desirable to
facilitate rapid and better germination.
 As per requirement of the crop the raised beds, furrows or
pits are prepared and field is kept ready for planting.
 In the riverbeds the trenches or pits are prepared as described
under muskmelon.

14. Sowing
 The sponge gourds are propagated through seeds.
 The seeds of sponge gourd are white and that of ridge gourd are
black.
 Seed rate for ridge gourd is 3.5-5.0 kg/ha and that of sponge
gourd is 2.5-3.5kg/ha.
 The seeds are sown in raised beds, furrows and pits.
 The seeds are soaked in water for 12-24 hours before sowing to
hasten the germination.

15. Spacing
 For sponge gourd the row to row distance of 1.5-2.5 m and hill
to hill distance of 60-120 cm are kept.
 Two seeds per hill in the case of raised beds and three to four
seeds per pit are sown.
 In Maharashtra, a spacing of (1.5x1.5) m is followed.
 With the pit system a spacing of 1.5-2.0 m between the two
rows and 1.0-1.5 m between the two plants is advocated.

16. Nutrition
 Farmyard manures at the rate of 15-20 tonnes /ha should be
mixed in the soil at the time of preparation of field.
 The super phosphate, potash and the half nitrogen should be
applied as a basal dose and one-half nitrogen should be given
as top dressing after one month of planting.
 Under Haryana conditions, farmyard manure 10-15 tonnes/ha
and 40-60kg N, 30-40kg P705 and 30kg of K,O/ha are
recommended for both the crops.
 In addition to N 20-30kg should be top dressed at the time of
vining.
 In Punjab, 100kg of N, 60kg of P205 and 60kg of K20/ha is
recommended.

17. Irrigation
 The summer crop must be irrigated regularly.
 The first irrigation should be given immediately after
the sowing. Summer crops require more frequent
irrigation than rainy season crops.
 Irrigation may be carried out according to the soil
moisture status and season.
 During the rainy season, irrigation may be required
during the early growth period.
 No systematic work is done on requirement of
irrigation water and its management in these crops.

18. Weed control
 Weeding should be done 15-20 days after the sowing.
 The second weeding may be done 20-25 days after the first one.
 The crop is kept weed free during the entire growth period.

19. Growth regulators
 In these crops, the female and male flowers are borne
separately on the same plant.
 The sex ratio can be regulated by exogenous application of
growth regulators NAA (200ppm) increases the female
flower production in ridge gourd and in turn increases the
yield significantly.
 In sponge gourd, exogenous application of Ethrel 250ppm
has been found to be beneficial.

20. Training and pruning
 In small homestead gardens, sponge gourd are trailed
over trelises arbours or pendals at 1.5-2.0m height.
 Commercial crop are trained on a kniffen system.
 The crop is trained when the seedlings are about 10-
15cm tall.
 The early crop can be allowed to trail on the bed
itself. In northern India the crop is generally not
trained and the vines are allowed to trail on the
ground.
 Rainy season crop should be staked so that the fruits
are prevented from direct contact with the soil.

21. Harvesting
 The crop is ready for harvest in about 60-90 days after the
seed sowing, depending upon the season and variety.
 The fruit attains a marketable maturity about 5-7 days after
anthesis of the female flower.
 Fruits should be harvested when they are still immature.
 Picking should be done every 3-4 days.
 If there is delay in harvesting, the fruits become more fibrous
and are unfit for human consumption.
 Fruits are harvested when they are tender and still immature.
 The flesh should not turn fibrous and picking should be done
earlier.
 The tenderness would decide edible maturity and not the fruit
size.

22. Yield
 Average yield of sponge is 8-12 tonnes/ha
 Hybrid: 20-25 tones/ha
 Grading according to the size
 packed in bamboo baskets with cushion
 Store in cool place for 3-4 days

23. Storage and marketing
 Harvesting should be done at proper edible maturity. Grading of fruits
should be done according to size.
 The fruits are packed in separate baskets. Care should be taken that the
fruits should not have any injury during transit.
 The fresh and tender fruits should reach the consumer. Fruits harvested at
the marketable stage can be stored for 3-4 days in a cool place without
any adverse effects.
 The fruits are packed in bamboo baskets with proper padding to prevent
injury in transit. The fruits are marketed in local and distant markets. The
plastic crates are also used for packing of fruits.

24. Research articles
 2.Mature Luffa Leaves (Luffa cylindrica L.pptx
 3.Selection Parameters in Sponge Gourd (Luffa
cylindrical.pptx

25. Mature Luffa Leaves (Luffa cylindrica L.) as a Tool for Gene
Expression Analysis by Agroinfiltration
Kamila Błaz˙ejewska1, Małgorzata Kapusta2, Elz˙bieta Zielin´ ska1, Zbigniew Tukaj1 andIzabela A. Chincinska1*
1 Department of Plant Physiology and Biotechnology, Faculty of Biology, University of Gdan´ sk, Gdan´ sk,
Poland,
2 Department of Plant Cytology and Embryology, Faculty of Biology, University of Gdan´ sk, Gdan´ sk, Poland

26. It can be used for genetic modification of mature plant tissues
To induce transient expression of genes in a plant
To date there are no literature sources on the possibility of transient
gene expression in mature cucurbit tissues
Agroinfiltration

27. 1. Comparison of cucurbit leaves absorption capacity.
2. To standardize the agroinfiltration protocol for generation of plant expression systems
based on mature cucurbit tissues.
3. To study the effect of agroinfiltration efficiency on the GUS activity in leaf blade and
exudates
OBJECTIVE OF THIS RESEARCH

28. Materials and Methods
For agroinfiltration analysis in luffa plants we used Agrobacterium tumefaciens
strain LBA 4404 transformed with a commercially available vector pRI 201-AN-
GUS containing b-glucuronidase gene (uidA) sequence under the control of the
CaMV 35S promoter.
The untransformed A. tumefaciens strain LBA 4404 was used as a negative control.

29. Materials and Methods
1. The leaves were treated with A. tumefaciens strain LBA 4404
transformed with the binary vector pRI 201-AN-GUS carrying E. coli
uidA gene encoding GUS.
2. As a control, we used luffa wild type (wt) leaves, untreated with the
bacteria
3. The leaves infiltrated with A. tumefaciens LBA 4404 without the
binary vector.

30. Materials and Methods
 The leaves were cut at the petiole bases, then transported to the
laboratory and stored in a closed container. High humidity in the
containers was held by placing wet paper towels at the bottom.
 Immediately before the infiltration the leaf was weighed, then placed
adaxial side down on a layer of soft paper towels designed to protect
the leaf from mechanical damage during the syringe infiltration.

32. Materials and Methods
Phloem Exudation
 After leaves were submersed under 2.5mM EDTA
solution (to prevent the contact with atmospheric
oxygen) and recut by 2–3 mm.
 The leaves were immediately transferred to dark
plastic tubes containing 5 mL of 2.5 mM EDTA
solution to facilitate exudation.
 To protect against evaporation of EDTA solution the
tubes were carefully sealed with parafilm.

33. Materials and Methods
Protein Analysis
• Leaves without petioles were ground in 30 mL of protein extraction buffer
[50 mM NaHPO4, pH 7.0, 10 Mm b-mercaptoethanol, 10 mM EDTA, 0.1%
(w/v) sodium lauryl sarcosine, and 0.1% (w/v) Triton X-100].
• The leaf extracts and the EDTA-exudates were used for the total soluble
protein (TSP) content measurement using the Bradford (1976) method and for
GUS activity assays.

34. Materials and Methods
Electrophoresis and Western Blotting
• SDS-PAGE was performed with acrylamide/bisacrylamide solution mix was in
ratio (29:1) in both stacking and resolving gels.
• The anti-b-glucuronidase (C-Terminal) primary antibody (Sigma–Aldrich) and
the goat anti-IgG rabbit coupled with HRP (horseradish peroxidase) secondary
antibody were used for GUS detection.
• A recombinant GUS protein from Escherichia coli (Sigma–Aldrich) was used
as a reference.

35. Materials and Methods
GUS Activity Analysis
• The histochemical GUS staining and fluorogenic GUS assays were
performed as described by Jefferson et al. (1987)
• Spectrophotometric and fluorescence measurements were done using
Varioscan Flash Multimode Reader (Thermo Scientific).

36. RESULTS
Luffa leaves was easily absorbed and quickly diffused in the adjacent tissue around the injection site

37. RESULTS-Differences in the leaf structure of cucurbit species.
Stained with Calcofluor White and propidium iodide
(A,B) Cucurbita pepo L. cv.
Makaronowa Warszawska.
(C,D) Cucurbita moschata Duch.
(E,F) Luffa cylindrica L.
(A,C,E) anatomical comparison of
cucurbit
leaves showed large air spaces in luffa
leaf mesophyll (E),
while the mesophyll cells in leaves of
both Cucurbita species were closely
packed(A,C).
ad D adaxial, ab D abaxial part of leaf in
(A) corresponds to all photos.
(B,D,F) stomata structure in cucurbit
leaves is similar in all species analyzed, but
the sub-stomatal air spaces (marked with an
arrow) is significantly greater in luffa
leaf (F) than in the both cucurbit (B,D).

38. RESULTS
The biomass of leaves was also measured up to three post-infiltration days to monitor a turgor loss in the leaves.
Demonstrates that the infiltration
procedures we used are insignificant
for the transpiration intensity in luffa
leaves

39. RESULTS The histochemical X-gluc staining of the GUS activity
in luffa leaves and EDTA-exudates
(A) Leaf disks (10 mm in diameter) randomly collected from:
the leaves infiltrated with A. tumefaciens carrying Pri 201-
AN-GUS, the leaves infiltrated with A. tumefaciens LBA
4404 without the binary vector, wt leaves.
(B) Petiole cross section of the leaf infiltrated with A. t
umefaciens carrying pRI 201-AN-GUS and from luffa wt
leaf. The blue stained vascular bundles are shown.
(C) GUS activity in EDTA-exudate samples detected with X-
gluc on microtiter plate. The 100 mL samples were
collected at 1 dpi from wt leaves (wells A1–A9 and B1–
B9), from the leaves infiltrated with untransformed A.
tumefaciens LBA 4404 (wells C1–C9 and D1–D9) and
from the leaves infiltrated with A. tumefaciens carrying
pRI 201-AN-GUS (wells C10–C12, D10–D12, E1–E12,
F1–F12, G1–G2, H1–H2).
The EDTA-exudates, increased with the increase of the agroinfiltration
efficiency.

40. RESULT- Effect of agroinfiltration efficiency on the
GUS activity in leaf blade and exudates.
To estimate the efficiency of bacteria
introduction a leaf weight increase after
infiltration was calculated as percentage values
relative to the weight before the infiltration.
We divided the infiltrated leaves (n D 14) into
four classes (<10, 11–20, 21–30, and >30%)
depending on the leaf weight (the X-axis).
GUS activity (the Y-axis) in the whole leaf
blades extracts as well as in the EDTA-exudate
samples were measured at 1 dpi and then
normalized relative to the 100 cm2 of leaf
surface area [mM 4-MU/min in 100 cm2].

41. CONCLUSION
• The susceptibility of their tissues to the introduction of bacterial suspension
• Only luffa leaves revealed an excellent absorption property.
• This absorption property allows the large volumes of liquid containing high
density suspensions of Agrobacterium cells.
• Loose structure of the mesophyll tissue with the large air spaces
• High humidity promotes stomatal opening, which in consequence, increases
access to the air spaces located in the deeper leaf tissues

42. POSSIBILITY
• Interesting to use different extracellular polysaccharides isolated from
suitable Pseudomonas species as adjuvants to support the penetration of
Agrobacterium into the interior of the leaves
• To use luffa leaves as a tool for studies requiring the application of liquids
into the leaf tissue.
• Especially for evaluating phloem functions in different plant species

43. Selection Parameters in Sponge Gourd (Luffa cylindrical
Roem.) for Yield and Yield Related Component Traits
VIVEK PANDEY*, V. B. SINGH1 AND M. K. SINGH2
Department of Vegetable Science,! Narendra Deva University of Agriculture & Technology
Kumarganj, Faizabad 224229, India
2 C. S. Azad University of Agriculture & Technology
Kanpur 208002, India
Environment & Ecology 30 (2) : 412—414, April —June 2012

44. INTRODUCTION
The main aim of any breeding programme is to search out high
yielding varieties along with greater fruit number, weight, uniform
thickness, cylindrical fruit, free from bitterness, high female : male
sex ratio, earliness, non-fibrous fruit at edible stage and resistance to
powdery mildew and downy mildew as required.

45. METHODS
• Thirty sponge gourd genotypes including one check variety i.e., Pusa
Chikani, evaluated in randomized block design (RBD) with 3 replication
• The observation was recorded on eleven characters
• The estimation of coefficient of variation, heritability (h2= σ2
g /σ2
p x 100)
and genetic advance (GA= K x h² x σp,) was made using the standard
formula

46. Source df Sum of squares Mean squares Expectations MS
Blocks/Replication r-1 - - -
Between genotypes t-1 SS1 MS1 σ2e + rσ2g
Within varieties or error (r-1)(t-1) SS2 MS2 σ2e
Total (rt-1)
ANOVA Table
Where r- Number of replications
t – number of genotypes/treatments

47. ANOVA
• From the above table, environmental, genotypic and
phenotypic variances are estimated as detailed below.
• Environmental variance = σ2e
• Genotypic variance (σ2g) = MS1-MS2/r
• Phenotypic variance (σ2p) = σ2g+ σ2e

48. • ANOVA also permits estimation of phenotypic, genotypic and environmental co-
efficients of variation.
• Phenotypic co-efficients of variation (PCV) = σp/Meanx100
• Genotypic co-efficients of variation (GCV) = σg/Mean x 100
• Environmental co-efficients of variation (ECV) = σe/Mean x 100
Where σp, σg, σe are phenotypic, genotypic and environmental standard deviations
respectively.
ANOVA

49. Results and Discussion
Analysis of variance for 11 characters in spongs gourd. * Significant at 5% probability
level, ** Significant at 1% probability level.
This suggested that the materials having adequate variability and response to selection may be expected in
the breeding program

50. Results and Discussion
Range, mean, coefficient of variation, heritability and genetic advance for different
character in sponge gourd
The wide differences between PCV and GCV implied their susceptibility to environmental fluctuation

51. CONCLUSION
Based on these study, it can be concluded that characters, average fruit
length, number of fruit per plant and average fruit weight exhibited
high PCV, GCV, heritability and genetic advance, indicated that the
selection for the above mentioned three characters, will be more
effective and reliable.

52. Thank you