ADVANCES IN CROP PRODUCTION AND IMPROVEMENT OF SPICE CROPS

1. WELCOME
TOPIC: ADVANCES IN CROP
PRODUCTION AND IMPROVEMENT
OF SPICE CROPS
LABIBA RIYAZ SHAH
SKUAST-K

2. SPICE: A spice is a seed, fruit, root, bark, or other
plant substance primarily used for
flavoring, coloring or preserving food.
A. Thapa et al.,2017

3. HISTORY OF
SPICES:
• The history of spices is almost as old as human
civilization.
• Spices were among the most valuable items of trade in
ancient and medieval times.
• As long ago as 3500 BC the ancient Egyptians were
using various spices for flavoring food, in cosmetics,
and for embalming their dead.
• The use of spices spread through the Middle East to
the eastern Mediterranean and Europe. Spices from
China, Indonesia, India, and Ceylon (now Sri Lanka)
were originally transported overland by donkey or
camel caravans.
• For almost 5000 years, Arab middlemen controlled
the spice trade, until European explorers discovered a
sea route to India and other spice producing countries
in the East.
• The ancient Indian epic Ramayana mentions cloves.

4. Important Landmarks in the History of Spices:
Pyramid age remarks about spices in Egyptian history and the use of spices in
'Mummies' as preservative.
6000-2100 BC
Vasco de Gama discovered the sea route to India and arrived at Calicut. 1498
Establishment of the British East India Company for trading in spices. British
landed in India on 24 August. 1600 at Surat.
1600
America entered the pepper trade 1795-1800
First research station for pepper established in India at Panniyur Kerala 1952-1953
Establishment of International Pepper Community at Jakarta (Indonesia) 1972
Establishment of National Research Centre for Spices (NRCS) 1986
NRCS Upgraded as Indian Institute of Spice Research. (IISR) 1996
Spices Board of India

5. Spice scenario
• India- Largest producer, consumer and exporter of spices
• China - second largest producer
• >90% production in India for domestic consumption
• Out of the 109 spices listed by the ISO, India produces as many as 75 in its various agro climatic
regions
• Throughout the world, and it has been estimated that spice crops are grown on an area of 8
million ha globally contributing to 31.6 million tons of spices annually.
• India’s share to world spices production is 6 million tons. The global spice industry amounts to
1.1 million metric tons, accounting to US$3.475 billion in value.
• India’s share at the global level is 0.575 million metric tons, accounting to US$2.037 billion, i.e.,
52 % in volume and 58.6 % in value
• Value-added spices (Curry powder/paste, mint products and spice oils and oleoresins) has
contributed around 43 per cent of the total foreign earnings during this period.
• Export of pepper was the major contributor, which had gone up to 18850 tonnes valued at Rs
7788.5 lakhs.
(Source: Spices Board of India 2014 )

6. Area and production statistics for different
spice crops in India:
SPICE AREA (ha) PRODUCTION (MT)
Chillies 721 1690
Clove 2 2
Coriander 468 567
Turmeric 246 931
Ginger 174 1846
Cardamom 78 22
Fennel 90 157
Black pepper 139 62
Fenugreek 122 189
NHB; 2019

7. STATE AREA (ha) PRODUCTION (MT)
Rajasthan 928.45 942.49
Madhya pradesh 640.17 2961.02
Gujrat 490.73 747.15
Uttar Pradesh 393.80 742.61
Andra Pradesh 199.05 742.61
Jammu and Kashmir 4.85 1.14
Area and production statistics for leading
spice producing states in India:
NHB; 2019

8. Importance and scope
• India has diverse soil and climate & several agro- ecological regions which
provides the opportunity to grow a variety of spice crops.
• Spices are low volume and high value crop.
• Spice crops play an important role in India’s economy by improving the
income of the rural people
• These crops are labor intensive so generate lot of employment opportunities
for the rural population.
• The increasing demand of Indian spices in foreign markets offers scope for
higher incomes.
• Spices can improve the palatability and the appeal of dull diets.
• Spices are used in pharmaceutical, cosmetic industries and also in religious
rituals.

9. Medicinal properties of few major spices
Chilli Digestive, thermogenic, carminative, stimulant, cardiotonic, antipyretic, serdorific, rubefacient & sialagogue. Caraway
Stomachic, carminative, anthemintic, lactagogue, adjuvant/ corrective for nauseating & griping effects of medicines
Cardamom Stimulant, tonic, diuretic, carminative, digestive, expectorant, cardiotonic & used in several pharmaceutical preparations.
Cardamom (large) Hypnotic, appetizer, astringent to bowels, tonic to heart and liver.
Corriander Carminative, diuretic, tonic, stimulant, stomachic, refrigerant, aphrodisiac, analgesic, anti-inflammatory Cumin Digestive,
carminative, astringent, anti-inflammatory, constipating, diuretic, revulsive, galactogogue, uterine & nerve stimulant
Pepper . Pepper Anthelmintic, carminative, alterant, antiperiodic, diuretic, digestive, emmenagogue, rubefacient, stimulant,
stomachic, used in fever, asthma, cough, dyspepsia, flatulence, arthritis. Pepper long Expectorant, thermogenic, diuretic,
tonic, purgative, stomachic, digestive, emollient, antiseptic, used in bronchitis, fever, asthma. Pomegranate Astringent,
cooling, tonic, aphrodisiac, laxative, diuretic, cardiotonic, used in pectoral diseases
Saffron Stimulant, tonic, stomachic, aphrodisiac, anodyne, anti spasmodic, emmenagogue, diuretic, laxative, used in bronchitis,
fever, epilepsy, skin diseases, decolouration of skin.
Turmeric Thermogenic, emollient, anodyne, anti inflammatory, vulnerary, depurative, antiseptic, appetizer, carminative,
expectorant, stomachic, anthelmintic, stimulant, ophthalmic, tonic, used in skin diseases, dyspepsia, asthma, cough,
bronchitis, inflammations, ulcers, worms, skin discolouration.
Ginger Digestive, carminative, emollient, appetizer, stomachic, rubefacient, anodyne, expectorant, anthelmintic, stimulant.
Greater galanga Carminative, expectorant, digestive, vulnerary, febrifuge, stimulant, depurative, used in skin diseases,
rheumatism, asthma, wounds, fever, haemorrhoids. Horse radish Thermogenic, appetizing, digestive, stomachic, laxative,
anti inflammatory, anodyne, refreshing, antibacterial. Hyssop Stimulant, carminative, pectoral, used in nervous disorders,
toothache, pulmonary & uterine troubles

10. Products obtained from various spices

11. Categories of spices
Major spices
Black pepper Chillies Ginger Turmeric

12. Categories of spices
Seed spices
Coriander Fennel Cumin

13. Categories of spices
Tree spices
Clove Nutmeg Kokum

14. Categories of spices
Miscellaneous
Garlic Saffron

15. ADVANCES IN SPICE
PRODUCTION AND
IMPROVEMENT:

16. Why need for improvement?
1. Ginger and turmeric normally propagates by rhizome with a low proliferation rate, and the reproducing part (rhizome) is also the
economically used part of the plant, which restricts the availability of ginger seeds needed for cultivation,
2. easily infected by soil-borne pathogens such as bacterial wilt (Pseudomonas solanacearum), soft rot (Pythium aphanidermatum)
and nematodes (Meloidogyne spp.), which cause heavy losses in yield,
3. Rhizomes show variations and degeneration under long-term vegetative propagation and
4. Normal breeding of vegetatively propagated plants is a real problem due to poor flowering and seeds set.
5, In turmeric planting material requirement is about 2.5 t/ha and total requirement of the country is about 2 lakh tonnes per year.
In addition, during storage and cultivation, rhizomes are susceptible to rhizome rot disease that causes tissue senescence and
degeneration. It is difficult to propagate through seed because of poor flowering and seed set. Survival percentage of plants through
rhizome is low (80 per cent) in field condition. The availability of quality planting material is also scarce during the cropping season
(June - September).
6. Black pepper can be propagated by seeds, cuttings, layering, and grafting however seed propagation often results in genetic
variation due to formation of recombinants while other methods of black pepper propagation are slow and time consuming (Atal &
Banga, 1962).
7. Saffron and garlic are sexually sterile. Saffron is propagated by corms as the flowers are sterile and fail to produce viable seeds. A
corm survives for only one season, producing up to ten “cormlets” that eventually give rise to new plants (Deo 2003).
8. Lack of high yielding cultivars.
9. Abiotic stresses e.g salt stress in cumin and moisture stress in saffron at the end of growth (Javed et al., Ascough et al. 2009)

17. PROPOGATION
METHODS AND
MICROPROPAGATION

18. TRENCH METHOD IN BLACK PEPPER PROPOGATION
In this method, a trench of 45 cm depth, 30 cm
width and of convenient length is made.
Split halves of bamboo or split halves of
PVC pipes are fixed at 45° angle by
keeping split portion facing upward
Rooted cuttings are planted in the trench at
the rate of one cutting for each bamboo split.
After ten days Each single nodded cutting with
the bunch of roots intact is cut and planted in
polythene bags filled with fumigated potting
mixture.
The buds start developing in about three weeks
and then the poly bags can then be removed and
kept in shade till main field planting. The
advantages of this method of propagation are
•rapid multiplication rate (1:40)
•well developed root system
•higher field establishment and
vigorous growth as a result of better root system

19. SERPENTINE METHOD
Three node cuttings planted in polythene bags.
The method requires grown up rooted cuttings
in 20 x 10 cm poly bags.
When the plant develops two leaves they are trailed
horizontally in polythene bags containing potting
mixture kept below each tender node. Each node will
be pressed into the mixture with polythene bags with
`V' shaped midribs of coconut leaves. As new shoots
arise these will be trailed horizontally in polythene
bags containing potting mixture
Once twenty nodes get rooted first 10 bags
in the rooted nodes will be separated by
cutting at the inter nodes. The inter nodal
stub will be pushed back into the potting
mixture. These stubs also produce a second
root system.
After three months it will be ready for planting in the main
field. On an average 60 cuttings will be obtained in a year by
this method from each mother cutting. Recommended in
black pepper nurseries for large scale multiplication.

20. The vines in soil mound
method exhibited superior
performance with respect to
length and root production
per node in all the year of
study.
The success of cuttings from
soil mound method was also
the highest.
The rooting percentage of
cuttings obtained from soil
mound and bamboo split
method were superior over
traditional method of
multiplication.
The B:C ratio for marketable
black pepper propagule by
soil mound method was the
best (2.13:1)
Explicating proper multiplication method for black pepper propagation:
GROWTH PERFORMANCE OF BLACK PEPPER VINES
UNDER DIFFERENT MULTIPLICATION METHODS
Treatment Length of
vine (m)
Node/vine
(no)
Roots/node(no) Branches/vine
(no)
T1 1.65 13.50 4.12 1.00
T2 2.10 17.60 4.56 2.30
T3 1.24 9.40 2.90 1.05
T4 _ _ _ _
SD 0.43 4.1 0.86 0.73
T1:BAMBOO SPLIT METHOD.,T2:SOIL MOUND METHOD.,T3:SERPENTINE METHOD.,T4:CONVENTIONAL METHOD

21. SINGLE BUD RHIZOME TECHNIQUE OF TURMERIC FOR
SEEDLINGS PRODUCTION IN PROTRAYS
Advantages:
1. Reduction in the requirement of
seed rhizome quantity by 25 per cent.
2. Saving huge quantity of rhizome
which can be used for commercial
purpose.
3. Reduction in the cost of planting
material.
4. Saving land usage 1-2 months from
normal duration period of the crop.
5. Overcoming the disease incidence
and also a screening for disease
infected materials
6. Overcoming the delay in monsoon
arrival up to 1-2 months
Malhotra et al.,2013

22. In vitro minirhizome production in turmeric (Curcuma longa L.) cultivar Alleppey
Supreme and its comparative anatomical and histochemical analysis:
This study was conducted to develop
an efficient protocol for the
development of microrhizome
production in Alleppey Supreme,
using two media combinations and
four types of culture vessels. It was
observed that the variety showed
highest response in liquid MS
medium with 80gl-1 sucrose in
Planton culture vessels. Yield was
nearly three times higher in
minirhizome seed material (526.67)
compared to the conventional
rhizome.
GROWTH PARAMETERS:
YIELD
Archana et al.,2014

23. ANATOMICALAND HISTOCHEMICAL COMPARISON OF RHIZOMES
Archana et al.,2014

24. In vitro propagation of ginger (Zingiber officinale)
An efficient and promising protocol for in
vitro propagation of Zingiber officinale
Rosco using sprouting buds was
established. Sprouting buds were
sterilized and cultured onto MS medium
supplemented with different growth
regulators. Augmentation of MS-medium
with 4.5 mg/l BAP recorded the highest
percentage of shootlets multiplication.
Shootlets were highly rooted on half
strength of B5 medium supplemented
with 1.0 mg/l NAA. The maximum
percentage of acclimatization, hardening
and rhizomes production of in vitro
derived plants in greenhouse was 80–
100%. Multiple shootlets formation/bud of Zingiber officinale after 4 weeks of cultivation on MS
medium supplemented with, (A) 4.5 mg/l BAP, (B) 4 mg/l Kin + 1 mg/l NAA, (C) 3.5 mg/l BAP
and (D) 1 mg/l BAP + 0.2 mg/l IAA.
Abbas et al.,2011

25. Rooting response of Z. officinale cultured on MS medium containing various
concentrations of NAA after 4 weeks of cultivation. Where (A) represent roots
formation per shoot, (B) 0.8 mg/l NAA, (C) 1 mg/l NAA and (D) 1.2 mg/l NAA.
In vitro well developed roots of Zingiber officinale prior transfer to field, (A)
half strength B5 medium, (B) half strength Heller’s medium, (C) half
strength NN medium and (D) half strength MS medium.
Type of nutrient media Roots number/shoot Root length (cm)
Half strength B5 25.3 7.93
Half strength Heller’s 23.7 7.33
Half strength NN 19.0 6.30
Half strength MS 12.3 3.60
Abbas et al.,2011

26. Acclimatization of micro propagated Zingiber officinale plantlets under
greenhouse conditions, (A) after 2 weeks of transferring and (B) 2 month old
plants, (C) after 7 months.
Abbas et al.,2011

27. Garlic shoot-tips were cultured on LS medium to
regenerate proliferative shoots. These shoot-tips
produced multiple shoots cultured at 20°C under
12-h light conditions. After 2-months culture under
25°C and 16-h light or 20°C and 12-h light, bulblets
with a weight of 30 to 300 mg were formed. The
frequency of bulblet formation was higher under
25°C and 16-h light. The early maturing cultivars,
such as cv. Isshu-wase, cv. Shannhai, cv. Isshu-
gokuwase and cv. Santo, formed bulblets at a rate of
37 to 74%, whereas the late maturing cultivars, such
as cv. Furano and cv. Howaito-roppen, formed
bulblets at a rate of 13 and 2%, respectively, during
2 months incubation. Effect of low temperature
treatment of germination of bulblets resulted in high
germination.
A novel micropropagation method for garlic (Allium sativum L.) by the combination of
initial shoot-tip culture, shoot multiplication and in vitro bulblet formation
Cultivar Culture condition
200c/12hr 250c/16hr
No of shoots
cultured
Bulblet
formation %
No of shoots
cultured
Bulblet
formation %
Isshu-wase 3 0 19 74
Shanhai 6 33 47 72
Ishu gokuwase 12 8 76 39
Santo 13 15 38 37
Furano 7 0 15 13
Howaito
roppen
7 0 43 2
Suma et al.,2008

28. Effects of high (35oc) middle (20oc) and low (50c)
temperatures on germination for 2 weeks.
(A-a) LS growth regulator-free medium, after 4 weeks of culture. (A-b) LS
NAA + BA medium (5 ~M NAA and 10 ~M BA) 4weeks of culture. (A-c) LS
IAA + BA medium (1 IxM IAA and 1 p,M BA), 4 weeks of culture. Bars = 1
mm. (B) Multiple shoots by shoot multiplication (C) In vitro bulblet
formation.

29. Various explants were cultured on
different nutrient media supplemented
with various concentrations of plant
growth regulators to standardize the
best media combination for obtaining
optimum response with respect to corm
production and development of Stigma
Like Structures (SLS). Highest
response (60%) was observed with half
ovaries on G-5 media supplemented
with 27μM NAA and 44.4μM BA.
Stigma Like Structures were developed
from half ovary explants both directly
and indirectly. Maximum number (120
indirectly and 20 directly) and size (5.2
cm) of SLS were obtained in G-5
medium supplemented with 27μM NAA
and 44.4μM BA.
Javid et al.,2011
In vitro development of microcorms and stigma like structures
in saffron (Crocus sativus L.)
Morphogenetic response of different explants cultured on G-5 medium supplemented with 3% sucrose.

30. In vitro development of micro corms and SLS in saffron, A: Shoot formation from
apical bud, B and C: Micro corms formation from apical bus, D: Indirect SLS
development from half ovary, E and F: Direct SLS development from half ovary.

31. GENETIC
TRANSFORMATION

32. Agrobacterium tumefaciens mediated transformation and
regeneration of ginger (Zingiber officinale Rosc.)
The absence of seed set in ginger makes conventional
breeding methods inapplicable warranting genetic
modification through biotechnological means.
Agrobacterium tumefaciens strain EHA105/p35SGUSInt,
effective in expressing b-glucuronidase activity, was
used to standardize the pre-culture of explants, bacterial
dilution, and co-cultivation period, besides evaluating
the effect of acetosyringone and post cultivation in
darkness, and to assess the optimum concentration of
kanamycin as selection agent for transformation.
Transformants were recovered on selection media
containing 100 mg L–1 kanamycin and a combination of
2,4- D 1.0 mg L–1 and BA 0.5 mg L–1, and regenerated in
half strength MS media of BA 3.0 mg L–1 and 2,4-D 0.5
mg L–1. Successful transformation was confirmed by
histochemical GUS assay and polymerase chain reaction
analysis.
Suma et al., 2008

33. Suma et al., 2008

34. Somatic
embryogenesis
and transgenic
development in
black pepper for
delayed infection
and decreased
spread of foot rot
caused by
Phytophthora
capsici
Somatic embryogenesis was induced successfully from tender leaf explants of in vitro
established mature tissues using the MS medium. Successful transformation of leaf explants
from these somatic embryo derived plants using Agrobacterium strain pGV 2260 carrying
‘osmotin’ under the control of CaMV 35S promoter was achieved. About 50 putative
transgenic plants were obtained and planted in the green house. The gene transfer was
confirmed by polymerase chain reaction (PCR) using osmotin and npt II specific primers.
Infection with P. capsici on detatched leaves of putative transgenics showed delayed infection
and decreased rate of disease spread indicating differential expression of osmotin. Stem and
leaf inoculated plants showed survival. It can be inferred that osmotin confers resistance to
infection by Phytophthora capsici in black pepper.
Sinoj et al., 2013

36. Development of CMV and TMV resistant
transgenic chili pepper:
• Both cucumber mosaic virus (CMV) and tobacco mosaic
virus (TMV) coat protein (CP) genes have been transferred
to chili pepper (Capsicum annuum var. Longunt) cultivar
8212 by a modified procedure of Agrobacterium
tumefaciens-mediated transformation using hypocotyl as
the explant. PCR analysis revealed the presence of both
CMV and TMV CP genes in at least 11 primary
transformants out of 49 kanamycin-resistant chili pepper
plants. Integration and expression of CMV CP and TMV
CP transgenes in one of the homozygous line, 16-13, were
confirmed by Southern blot, RT-PCR and western blot
analyses. Line 16-13 was highly resistant to infection of
homologous CMV and TMV strains in greenhouse
conditions when successively challenged with CMV and
TMV or challenged with TMV alone. Furthermore, field
trials on T2, T3 and T4 progenies of Line 16-13 were
performed on scales of 123, 300 and 10,000 plants,
respectively, in consecutive years with the permission of the
Chinese government authority. The transgenic plants
displayed delayed symptom development and significantly
milder disease severity in field conditions when compared
to untransformed chili pepper plants, resulting in 47 and
110% increase in pepper fruit yield in surveys conducted in
respectively.
Cai et al., 2003

37. Cai et al., 2003

38. Somatic hybridization

39. Regeneration of somatic hybrids of ginger via
chemical protoplast fusion
Ginger (Zingiber officinale Rosc.) somatic
hybridization was attempted by using
polyethylene glycol (PEG)-mediated
protoplast fusion. Protoplasts of three
ginger cultivars isolated from the
embryogenic cell suspensions were fused
with each other. The highest binary fusion
rate [13.5% in the fusion of ginger ‘Lushan
Zhangliang jiang’ and ‘Chenggu Huang
Jiang’ was observed with the treatment of
30% PEG6000 and could regenerate
plantlets. Approximately 15 months were
used for the regeneration of whole plants,
and 15 shoots were obtained from the
fusion. Three plantlets were identified as
hybrids by using RAPD, and they were all
diploids by analysis with flow cytometry.
Guan et al., 2010

40. Guan et al., 2010

41. The SbNHX1 gene, cloned from an extreme halophyte Salicornia
brachiata was transformed in cumin using optimized in
planta transformation method. The SbNHX1 gene encodes a
vacuolar Na+/H+ antiporter and is involved in the
compartmentalization of excess Na+ ions into the vacuole and
maintenance of ion homeostasis Transgenic cumin plants were
confirmed by PCR using gene (SbNHX1, uidA and hptII) specific
primers. The single gene integration event and overexpression of
the gene were confirmed by Southern hybridization and
competitive RT-PCR, respectively. Transgenic lines L3 and L13
showed high expression of the SbNHX1 gene compared to L6
whereas moderate expression was detected in L5 and L10
transgenic lines. Transgenic lines (L3, L5, L10 and L13),
overexpressing the SbNHX1 gene, showed higher photosynthetic
pigments (chlorophyll a, b and carotenoid), and lower electrolytic
leakage, lipid peroxidation (MDA content) and proline content as
compared to wild type plants under salinity stress. Though
transgenic lines were also affected by salinity stress but performed
better compared to WT plants. The ectopic expression of
the SbNHX1 gene confirmed enhanced salinity stress tolerance in
cumin as compared to wild type plants under stress condition.
Schematic representation of optimized in planta transformation protocol used for
Agrobacterium-mediated genetic transformation of cumin seeds.
Sonika et al., 2016
In planta Transformed Cumin (Cuminum cyminum L.) Plants, Overexpressing the SbNHX1 Gene Showed
Enhanced Salt Endurance

42. In planta transformation of cumin seeds. Molecular confirmation of in planta transformed transgenic lines.

43. The phytohormone ethylene is involved in many
developmental processes, including leaf and flower
senescence. Ethylene is perceived by plants through
receptors that trigger the downstream signal
transduction pathway. The mutated ethylene receptor
ERS1 (ethylene response sensor) from Arabidopsis is
of a dominant negative nature and confers ethylene
insensitivity in Arabidopsis. To investigate if the altered
ERS1 gene can affect the tissue senescence in
heterologous plants, we introduced it into coriander by
Agrobacterium-mediated transformation. Transgenic
plants were regenerated by co-cultivating hypocotyl
segments with A. tumefaciens harboring binary vector
pCGN1547 that carried the ERS1 gene. The presence
and expression of the transgene were confirmed by
genomic Southern blot and reverse transcriptase- PCR
analyses. Leaf and flower senescence were delayed
significantly in the transgenic plants. The ability of the
mutated ERS1 gene to confer the ethylene insensitive
phenotype can be exploited for extending the shelf-life
Heterologous expression of Arabidopsis ERS1 causes delayed senescence in
coriander
Wang, 2016

44. Molecular analysis of the ERS1 gene in coriander. A Southern
blot analysis of EcoRV-digested genomic DNA from leaves of a
wild-type plant and putative transformants (lines 11, 17, 19, 21,
23). B RT-PCR analysis of the expression of ERS1 transgene in
selected lines of transformants. Two bands of sizes 540 bp and
370 bp are visible. The 370-bp band corresponds to the expected
fragment of the transcript (the difference between the gene copy
in the plasmid and this band is due to the existence of two
introns). Ubiquitin primers were used as a control, generating
the expected pattern of four bands in each lane, which indicates
uniform amplification in the PCR reaction. M Marker, pC
pCGN1547 plasmid control. C RT-PCR analysis of the
expression of native ACC oxidase (ACO) and ERS1 homologs
in selected transgenic coriander lines (transgenic line number is
indicated above each lane)
Wang, 2016

45. CONCLUSION:
1. A huge gap between demand and production demands more
scope for production and improvement.
2. Modern technologies like marker assisted selection, CRISPR-
CAS9, somatic hybridization etc can be used for high yielding
varieties.
3. Less work is done on improvement of spices despite being high
value crops therefore require more attention.
4. Introduction into new temperature regimes can be done.
5. More focus should be given on exploitation of medicinal
properties.