USPP22593P3 - Stevia plant named ‘T60’ - Google Patents

Stevia plant named ‘T60’ Download PDF

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USPP22593P3
USPP22593P3 US12/799,668 US79966810V USPP22593P3 US PP22593 P3 USPP22593 P3 US PP22593P3 US 79966810 V US79966810 V US 79966810V US PP22593 P3 USPP22593 P3 US PP22593P3
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stevia
variety
lines
reb
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Grania Viorela Garnighian
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Sweet Green Fields International Co Ltd
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SGF Holdings LLC
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
    • A01H5/02—Flowers
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/14—Asteraceae or Compositae, e.g. safflower, sunflower, artichoke or lettuce
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/14—Asteraceae or Compositae, e.g. safflower, sunflower, artichoke or lettuce
    • A01H6/1488—Stevia

Definitions

  • Botanical/commercial classification Stevia rebaudiana L.
  • Stevia rebaudiana is a shrubby perennial, growing up to 65-100 cm tall. It is a member of the Compositae family and originates from Paraguay. This species is grown commercially in Brazil, China, Paraguay, the United States, and other countries. The plants are grown so that sweet glycosides present in the leaves can be extracted for use as sweeteners. The sweetest of these glycosides is rebaudioside A (Reb A).
  • the development of new varieties of Stevia rebaudiana with high levels of Reb A is desirable.
  • the new Stevia variety ‘T60’ described herein is such a variety.
  • the new ‘T60’ variety was asexually reproduced from callus through an in vitro tissue culture technique performed in Woodland, Calif.
  • the major characteristics exhibited by the new Stevia variety ‘T60’ that distinguish it from other varieties include a high concentration of total glycoside and high concentration of Reb A in the leaves from total glycoside.
  • Other major characteristics that also distinguish the new Stevia variety ‘T60’ from other varieties include a tall, bushy stature that is resistant to high winds; spatulate-oblanceolate to ovate leaves; high yield foliage production. For example, ‘T60’ produced over 3000 lbs/acre in one harvest.
  • Still other major characteristics that distinguish the new Stevia variety ‘T60’ from other varieties include being virus free, resistant to Fusarium ssp. and Sclerotinia ssp., drought resistant, and resistant to compact argilous soils.
  • Another major characteristic exhibited by the new Stevia variety ‘T60’ that distinguishes it from other varieties is an average biological cycle of 90-115 days between 20-27 N,S Latt, giving the option of secondary harvest per annual cycle.
  • FIG. 1 depicts a color photograph of a fully mature ‘T60’ stevia plant near the end of the growing season.
  • Table 1 depicts the glycoside and Reb A percentages for the ‘Criola’ lines described above.
  • the four ‘Criola’ lines were planted in vitro in tissue culture medium.
  • the tissue culture material was multiplied by successively transferring plantlets in hormone-supplemented culture. Plantlets of each variety were planted into 2 inch plugs and then into 4 inch pots between December of year 1 and January of year 2. The plantlets were planted in a field in Woodland, Calif. between March and April of year 2. The plantlets were then allowed to flower, which occurred between July and August of year 2.
  • anthers from the flowered plantlet were collected before anthesis. Approximately 200 anthers were collected from plantlets of each of the four ‘Criola’ lines. The anthers were then plated in vitro to derive haploid lines. The in vitro tissue culture medium induced callus formation from the plated anthers. Callus formation was observed in October of year 2. Haploid lines were created from the formed callus. The process of creating elite Stevia lines required that the chromosome count of each haploid line be doubled, since haploid lines cannot survive as plants. The full complement of chromosomes was regenerated by using successive colchicine treatments of 20 ppm, 40 ppm, and 60 ppm on both the callus and the shoots regenerated from the callus.
  • Every green callus that formed a diploid plantlet was transferred into a hormone-free medium for multiplication. Multiplication of the diploid plantlets resulted in the creation of a unique line of plants with unique characteristics distinguishable form the original four ‘Criola’ lines.
  • These new Stevia lines were named T-lines.
  • the T-lines were grown out in 2 inch plugs, then transferred to 4 inch pots, and then planted in the field in Woodland, Calif.
  • the selected T-lines were shown to be superior to the parental stock.
  • the selected T-lines either had higher total levels of glycoside, or higher levels of Reb A. Based on these characteristics, all callus material that gave rise to the new T-lines was collected. Genetically unique lines were developed from this callus material.
  • the new T-lines were planted from tissue culture into 2 inch plugs and then transplanted into 4 inch pots. By the age of six weeks, this second generation of T-line plants had grown to about 10 inches in height in the pots. At six weeks the T-line plants were sampled to obtain HPLC data on the percentage of Reb A production. This was done even though the percentage of glycosides found in the leaf material of the new T-lines plants would not be as high as expected levels in mature plants. The levels of Reb A in these new T-line Stevia plants were as high as 89.2% of total glycoside levels. After the HPLC data was obtained, the T-line Stevia plants were planted in the field in Woodland, Calif., and grown to maturity. At maturity, the production rates of glycosides were evaluated.
  • the present Stevia variety ‘T60’ is one of the three selected T-lines developed by the methods described herein.
  • the Stevia variety ‘T60’ was derived from the T-line ‘T6’.
  • the ‘T6’ line was derived from ‘Criola’ line ‘IAN-135’.
  • Table 2 depicts HPLC data on the percentage of glycoside and Reb A for samples of the Stevia variety ‘T60’.
  • the HPLC data shows that the average percentage of Reb A to Glycoside in Stevia variety ‘T60’ was 83.2%. This is an increase of at least 11.5% over the ‘Criola’ lines. The ‘T60’ variety also showed an increase of 34.5% over ‘IAN-135’, from which ‘T60’ is derived.
  • the Stevia variety ‘T60’ is a perennial herb with filiform deep roots.
  • the ‘T60’ variety has a long stem that is on average 65-100 cm tall, erect, hairy, and slender.
  • the stem easily produces secondary shoots (suckers) from its base, dies off and is renewed annually.
  • the stem produces an average of 5 to 10 suckers.
  • the stem also produces numerous lateral branches, thus, forming a more-or-less roundish and dense crown.
  • the ‘T60’ variety also has an average biological cycle of 90-115 days. The variety goes dormant and is renewed annually.
  • the stem of the ‘T60’ variety produces an average of 3 to 7 lateral branches.
  • the average length of each lateral branches ranges from 30 to 40 cm.
  • the average diameter of the lateral branches ranges from 5 to 7 mm.
  • the lateral branches are light green in color (RHS 142A).
  • the lateral branches form a roundish and dense crown.
  • the crushed leaves exude a strong odor, and all the green parts of the plant taste sweet.
  • the leaves are simple, opposite and sub sessile.
  • the internodes have an average length of 2 to 4 cm.
  • the blades are subcoriaceous, very variable in shape and size, and ovate.
  • the blades have an average length of 2 to 3 cm and an average width of 0.6 to 1 cm.
  • the blades are apex obtuse, base cuneate, margins entire serate on the upper half, three primary veins arise from the leaf base raised and prominent on the blade's lower side, immersed on the upper side, and secondary venation reticulate.
  • the leaf blades of actively growing plants of the ‘T60’ variety are light green in color (RHS 142A). In the dry state, the leaf blades are olive-green to brownish green in color (RHS 147A-147B), and usually darker on the upper side. Both surfaces are subscabrous with black glandular dots on the lower side, the leaves sub sessile or the petiole to 3-4 mm long.
  • the capitula are arranged into loose, paniculate corymbose inflorescences at the terminal ends of the branches, and have a peduncle that is on average 1-4 cm long, and very slender.
  • each capitulum Pedicels of each capitulum are slender and on average are 1-4 mm long, bracts are linear-lanceolate, and on average are 1-2 mm long.
  • Each capitulum is enveloped by an involucre, and the lower half has a light green color (RHS 142A), and the upper half has a yellowish (RHS 142C) color.
  • the stems at maturity are brown-reddish in color (RHS 35A-35B).
  • the Stevia variety ‘T60’ has 5 phyllaries that are finely hairy, green (RHS 129B) when fresh, linear to subulate, 4-5 mm long, and acute to rounded at apex. Each capitulum is made up to 5 disk florets greenish white (RHS 155C). The seeds are achenes bearing numerous, equally long pappus awns.
  • the Stevia variety ‘T60’ expresses high concentrations of total glycosides, with a total of rebaudioside A concentration of greater than 80%.
  • the Stevia variety ‘T60’ has an average biological cycle of 90-115 days.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Physiology (AREA)
  • Botany (AREA)
  • Developmental Biology & Embryology (AREA)
  • Environmental Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

A new and distinct Stevia rebaudiana plant named ‘T60’ is described. The new variety is distinguished from other Stevia varieties by a high leaf concentration of rebaudioside A; its tall, bushy nature; its ovate leaves; and its long stem stature.

Description

Botanical/commercial classification: Stevia rebaudiana L.
Variety denomination: ‘T60’.
BACKGROUND OF THE INVENTION
Stevia rebaudiana is a shrubby perennial, growing up to 65-100 cm tall. It is a member of the Compositae family and originates from Paraguay. This species is grown commercially in Brazil, China, Paraguay, the United States, and other countries. The plants are grown so that sweet glycosides present in the leaves can be extracted for use as sweeteners. The sweetest of these glycosides is rebaudioside A (Reb A).
The development of new varieties of Stevia rebaudiana with high levels of Reb A is desirable. The new Stevia variety ‘T60’ described herein is such a variety.
The new ‘T60’ variety was asexually reproduced from callus through an in vitro tissue culture technique performed in Woodland, Calif.
BRIEF SUMMARY OF THE INVENTION
The major characteristics exhibited by the new Stevia variety ‘T60’ that distinguish it from other varieties include a high concentration of total glycoside and high concentration of Reb A in the leaves from total glycoside. Other major characteristics that also distinguish the new Stevia variety ‘T60’ from other varieties include a tall, bushy stature that is resistant to high winds; spatulate-oblanceolate to ovate leaves; high yield foliage production. For example, ‘T60’ produced over 3000 lbs/acre in one harvest. Still other major characteristics that distinguish the new Stevia variety ‘T60’ from other varieties include being virus free, resistant to Fusarium ssp. and Sclerotinia ssp., drought resistant, and resistant to compact argilous soils. Another major characteristic exhibited by the new Stevia variety ‘T60’ that distinguishes it from other varieties is an average biological cycle of 90-115 days between 20-27 N,S Latt, giving the option of secondary harvest per annual cycle.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 depicts a color photograph of a fully mature ‘T60’ stevia plant near the end of the growing season.
 DETAILED DESCRIPTION OF THE INVENTION
Four wild varieties of Stevia were imported from the Paraguayan government. These varieties were derived from a wild germ plasm in Paraguay, locally identified as ‘Criola’ material, and released by the Paraguayan government upon request for research and breeding. The four imported ‘Criola’ lines were named ‘IAN-135’, ‘IAN-142’, ‘IAN-228’, and ‘IAN-259’.
Table 1 depicts the glycoside and Reb A percentages for the ‘Criola’ lines described above.
TABLE 1
Plant Length of
Flowering Height Leaves
Variety Cycle Days (cm) (cm)
IAN-228 Early 110 65 4
IAN-135 Medium 180 82 4
IAN-259 Medium 180 55 5
IAN-142 Late 200 110 6
Width of
Leaves Total Total Reb A to
Variety (cm) Glycoside Reb A Glycoside
IAN-228 2 18.47 13.25 71.7%
IAN-135 2 16.56 8.06 48.7%
IAN-259 2 16.55 11.24 67.9%
IAN-142 3 19.06 13.15   69%
As a first step in the production of new Stevia varieties, the four ‘Criola’ lines were planted in vitro in tissue culture medium. The tissue culture material was multiplied by successively transferring plantlets in hormone-supplemented culture. Plantlets of each variety were planted into 2 inch plugs and then into 4 inch pots between December of year 1 and January of year 2. The plantlets were planted in a field in Woodland, Calif. between March and April of year 2. The plantlets were then allowed to flower, which occurred between July and August of year 2.
In August of year 2, anthers from the flowered plantlet were collected before anthesis. Approximately 200 anthers were collected from plantlets of each of the four ‘Criola’ lines. The anthers were then plated in vitro to derive haploid lines. The in vitro tissue culture medium induced callus formation from the plated anthers. Callus formation was observed in October of year 2. Haploid lines were created from the formed callus. The process of creating elite Stevia lines required that the chromosome count of each haploid line be doubled, since haploid lines cannot survive as plants. The full complement of chromosomes was regenerated by using successive colchicine treatments of 20 ppm, 40 ppm, and 60 ppm on both the callus and the shoots regenerated from the callus.
Every green callus that formed a diploid plantlet was transferred into a hormone-free medium for multiplication. Multiplication of the diploid plantlets resulted in the creation of a unique line of plants with unique characteristics distinguishable form the original four ‘Criola’ lines. These new Stevia lines were named T-lines. The T-lines were grown out in 2 inch plugs, then transferred to 4 inch pots, and then planted in the field in Woodland, Calif. The selected T-lines were shown to be superior to the parental stock. The selected T-lines either had higher total levels of glycoside, or higher levels of Reb A. Based on these characteristics, all callus material that gave rise to the new T-lines was collected. Genetically unique lines were developed from this callus material.
To test for percentage of Reb A production, the new T-lines were planted from tissue culture into 2 inch plugs and then transplanted into 4 inch pots. By the age of six weeks, this second generation of T-line plants had grown to about 10 inches in height in the pots. At six weeks the T-line plants were sampled to obtain HPLC data on the percentage of Reb A production. This was done even though the percentage of glycosides found in the leaf material of the new T-lines plants would not be as high as expected levels in mature plants. The levels of Reb A in these new T-line Stevia plants were as high as 89.2% of total glycoside levels. After the HPLC data was obtained, the T-line Stevia plants were planted in the field in Woodland, Calif., and grown to maturity. At maturity, the production rates of glycosides were evaluated.
All new T-lines were grown out in greenhouses in Woodland, Calif. for planting in the field, to allow for further testing and breeding for seed.
Of the new Stevia T-lines, three varieties were selected that produced high levels of Reb A. The production of high levels of Reb A in these Stevia T-lines will allow large amounts of Reb A to be harvested. The present Stevia variety ‘T60’ is one of the three selected T-lines developed by the methods described herein. The Stevia variety ‘T60’ was derived from the T-line ‘T6’. The ‘T6’ line was derived from ‘Criola’ line ‘IAN-135’.
Table 2 depicts HPLC data on the percentage of glycoside and Reb A for samples of the Stevia variety ‘T60’.
TABLE 2
Samples of mature Total Total Reb A to
‘T 60’ Stevia plants Glycoside Reb A Glycoside
09-0144 15.18 12.58 82.8%
09-0154 16.96 14.39 84.8%
09-0165 14.58 12.00 82.3%
09-0166 15.05 12.46 82.8%
The HPLC data shows that the average percentage of Reb A to Glycoside in Stevia variety ‘T60’ was 83.2%. This is an increase of at least 11.5% over the ‘Criola’ lines. The ‘T60’ variety also showed an increase of 34.5% over ‘IAN-135’, from which ‘T60’ is derived.
DETAILED BOTANICAL DESCRIPTION
The Stevia variety ‘T60’ is a perennial herb with filiform deep roots. The ‘T60’ variety has a long stem that is on average 65-100 cm tall, erect, hairy, and slender. The stem easily produces secondary shoots (suckers) from its base, dies off and is renewed annually. The stem produces an average of 5 to 10 suckers. The stem also produces numerous lateral branches, thus, forming a more-or-less roundish and dense crown. The ‘T60’ variety also has an average biological cycle of 90-115 days. The variety goes dormant and is renewed annually.
The stem of the ‘T60’ variety produces an average of 3 to 7 lateral branches. The average length of each lateral branches ranges from 30 to 40 cm. The average diameter of the lateral branches ranges from 5 to 7 mm. The lateral branches are light green in color (RHS 142A). The lateral branches form a roundish and dense crown.
The crushed leaves exude a strong odor, and all the green parts of the plant taste sweet. The leaves are simple, opposite and sub sessile. The internodes have an average length of 2 to 4 cm. The blades are subcoriaceous, very variable in shape and size, and ovate. The blades have an average length of 2 to 3 cm and an average width of 0.6 to 1 cm. The blades are apex obtuse, base cuneate, margins entire serate on the upper half, three primary veins arise from the leaf base raised and prominent on the blade's lower side, immersed on the upper side, and secondary venation reticulate.
The leaf blades of actively growing plants of the ‘T60’ variety are light green in color (RHS 142A). In the dry state, the leaf blades are olive-green to brownish green in color (RHS 147A-147B), and usually darker on the upper side. Both surfaces are subscabrous with black glandular dots on the lower side, the leaves sub sessile or the petiole to 3-4 mm long. The capitula are arranged into loose, paniculate corymbose inflorescences at the terminal ends of the branches, and have a peduncle that is on average 1-4 cm long, and very slender. Pedicels of each capitulum are slender and on average are 1-4 mm long, bracts are linear-lanceolate, and on average are 1-2 mm long. Each capitulum is enveloped by an involucre, and the lower half has a light green color (RHS 142A), and the upper half has a yellowish (RHS 142C) color. The stems at maturity are brown-reddish in color (RHS 35A-35B).
The Stevia variety ‘T60’ has 5 phyllaries that are finely hairy, green (RHS 129B) when fresh, linear to subulate, 4-5 mm long, and acute to rounded at apex. Each capitulum is made up to 5 disk florets greenish white (RHS 155C). The seeds are achenes bearing numerous, equally long pappus awns.
The Stevia variety ‘T60’ expresses high concentrations of total glycosides, with a total of rebaudioside A concentration of greater than 80%. The Stevia variety ‘T60’ has an average biological cycle of 90-115 days.

Claims (1)

We claim:
1. A novel and distinct variety of Stevia plant named ‘T60’ having the characteristics described and illustrated herein.
US12/799,668 2010-04-28 2010-04-28 Stevia plant named ‘T60’ Active USPP22593P3 (en)

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Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20110111115A1 (en) * 2009-11-06 2011-05-12 Jingang Shi Rebaudioside a polymorphs and methods to prepare them
US9131718B2 (en) 2009-06-16 2015-09-15 Epc (Beijing) Natural Products Co., Ltd. Process for rebaudioside D
US9578895B2 (en) 2010-08-23 2017-02-28 Epc (Beijing) Natural Products Co., Ltd. Rebaudioside A and stevioside compositions
USPP27815P3 (en) 2014-12-08 2017-03-28 S&W Seed Company Stevia plant named ‘SW 201’
USPP27937P3 (en) 2014-11-30 2017-04-25 S&W Seed Company Stevia plant named ‘SW 107’
USPP28373P3 (en) 2015-11-17 2017-09-12 S&W Seed Company Stevia plant named ‘SW 129’
US9795156B2 (en) 2011-03-17 2017-10-24 E.P.C (Beijing) Plant Pharmaceutical Technology Co., Ltd Rebaudioside B and derivatives
USPP28977P3 (en) 2016-03-31 2018-02-20 S&W Seed Company Stevia plant named ‘SW 227’
US10264811B2 (en) 2014-05-19 2019-04-23 Epc Natural Products Co., Ltd. Stevia sweetener with improved solubility
US10357052B2 (en) 2014-06-16 2019-07-23 Sweet Green Fields USA LLC Rebaudioside A and stevioside with improved solubilities
US10485256B2 (en) 2014-06-20 2019-11-26 Sweet Green Fields International Co., Limited Stevia sweetener with improved solubility with a cyclodextrin

Families Citing this family (1)

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US20160338396A1 (en) * 2010-12-31 2016-11-24 Glg Life Tech Corporation High Rebaudioside-A Plant and Methods of Producing the Same and Uses Thereof

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9131718B2 (en) 2009-06-16 2015-09-15 Epc (Beijing) Natural Products Co., Ltd. Process for rebaudioside D
US9635878B2 (en) 2009-06-16 2017-05-02 Epc (Beijing) Natural Products Co., Ltd. Process for rebaudioside D
US20110111115A1 (en) * 2009-11-06 2011-05-12 Jingang Shi Rebaudioside a polymorphs and methods to prepare them
US11202462B2 (en) 2010-08-23 2021-12-21 Sweet Green Fields International Co., Limited Rebaudioside A and stevioside compositions
US9578895B2 (en) 2010-08-23 2017-02-28 Epc (Beijing) Natural Products Co., Ltd. Rebaudioside A and stevioside compositions
US10285425B2 (en) 2010-08-23 2019-05-14 Epc Natural Products Co. Ltd Rebaudioside A and stevioside compositions
US11510428B2 (en) 2011-03-17 2022-11-29 Sweet Green Fields International Co., Limited Rebaudioside B and derivatives
US9795156B2 (en) 2011-03-17 2017-10-24 E.P.C (Beijing) Plant Pharmaceutical Technology Co., Ltd Rebaudioside B and derivatives
US10264811B2 (en) 2014-05-19 2019-04-23 Epc Natural Products Co., Ltd. Stevia sweetener with improved solubility
US11206857B2 (en) 2014-05-19 2021-12-28 Sweet Green Fields International Co., Limited Stevia sweetener with improved solubility
US11241031B2 (en) 2014-06-16 2022-02-08 Sweet Green Fields Usa, Llc Rebaudioside A and stevioside with improved solubilities
US10357052B2 (en) 2014-06-16 2019-07-23 Sweet Green Fields USA LLC Rebaudioside A and stevioside with improved solubilities
US10568351B2 (en) 2014-06-16 2020-02-25 Sweet Green Fields USA LLC Rebaudioside A and stevioside with improved solubilities
US10485256B2 (en) 2014-06-20 2019-11-26 Sweet Green Fields International Co., Limited Stevia sweetener with improved solubility with a cyclodextrin
USPP27937P3 (en) 2014-11-30 2017-04-25 S&W Seed Company Stevia plant named ‘SW 107’
USPP27815P3 (en) 2014-12-08 2017-03-28 S&W Seed Company Stevia plant named ‘SW 201’
USPP28373P3 (en) 2015-11-17 2017-09-12 S&W Seed Company Stevia plant named ‘SW 129’
USPP28977P3 (en) 2016-03-31 2018-02-20 S&W Seed Company Stevia plant named ‘SW 227’

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