USO0PP22593P3

(12) United States Plant Patent

(10) Patent No.:

Garnighian

(45) Date of Patent:

(54) STEVIA PLANT NAMED ‘T60’ (50)

Latin Name:

(51)

Stevia rebaudiana L.

Inventor:

58

Grania Viorela Garnighian, Prior Lake,

(

Mar. 20, 2012

Int. Cl. A01H 5/00

Varietal Denomination;

(75)

US PP22,593 P3

(200601)

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See application ?le for complete search history.

(73) Assignee: SGF Holdings LLC, Bellingham, WA (*)

Notice:

(Us)

Primary Examiner * Susan McCormick Ewoldt

Subject to any disclaimer, the term of this patent is extended or adjusted under 35

(74) Attorney’ Agent’ or Firm * Morrison & Foerster LLP

U.S.C. 154(1)) by 0 days.

(57)

ABSTRACT

(21) APPI' NO‘: 12/799,668

A new and d1st1nct Sleyza rebourlzano plant named T60 is

(22) Filed:

varieties by a high leaf concentration of rebaudioside A; its tall, bushy nature; its ovate leaves; and its long stem stature.

described. The new variety is distmguished from other Slevza

Apr. 28, 2010

(65)

Prior Publication Data US 2011/0271413 P1

1 Drawing Sheet

Nov. 3, 2011

1

2

Botanical/commercial classi?cation: Slevia rebaudiana L.

DETAILED DESCRIPTION OF THE INVENTION

Variety denomination: ‘T60’. Four wild varieties of Slevia were imported from the Para

BACKGROUND OF THE INVENTION

guayan government. These varieties were derived from a wild

germ plasm in Paraguay, locally identi?ed as ‘Criola’ mate rial, and released by the Paraguayan government upon request for research and breeding. The four imported ‘Criola’

Slevia 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 commer

lines were named ‘IAN-135’, ‘IAN-142’, ‘IAN-228’, and

cially in Brazil, China, Paraguay, the United States, and other

‘IAN-25 9’ .

countries. The plants are grown so that sweet glycosides present in the leaves can be extracted for use as sweeteners. 10

The development of new varieties of Slevia rebaudiana with high levels of Reb A is desirable. The new Slevia 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

TABLE 1

Variety

in Woodland, Calif. BRIEF SUMMARY OF THE INVENTION

The major characteristics exhibited by the new Slevia vari ety ‘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 char acteristics that also distinguish the new Slevia variety ‘T60’ from other varieties include a tall, bushy stature that is resis tant to high winds; spatulate-oblanceolate to ovate leaves;

20

25

high yield foliage production. For example, ‘T60’ produced over 3000 lbs/acre in one harvest. Still other major character

istics that distinguish the new Slevia variety ‘T60’ from other varieties include being virus free, resistant to Fusarium ssp. and Sclerolinia ssp., drought resistant, and resistant to com

between 20-27 N,S Latt, giving the option of secondary har

FIG. 1 depicts a color photograph of a fully mature ‘T60’ slevia plant near the end of the growing season.

Length of

Height

Leaves

Days

(cm)

(cm)

IAN-22 8

Early

110

65

4

IAN- 135 IAN-259 IAN- 142

Medium Medium Late

180 180 200

82 55 110

4 5 6

Width of Leaves

Total

Total

Reb A to

Variety

(cm)

Glycoside

Reb A

Glycoside

IAN-228 IAN-135 IAN-259 IAN-142

2 2 2 3

18.47 16.56 16.55 19.06

13.25 8.06 11.24 13.15

71.7% 48.7% 67.9% 69%

cessively transferring plantlets in hormone-supplemented

the new Slevia variety ‘T60’ that distinguishes it from other varieties is an average biological cycle of 90-115 days

BRIEF DESCRIPTION OF THE DRAWINGS

Cycle

Plant

Flowering

As a ?rst step in the production of new Slevia varieties, the four ‘Criola’ lines were planted in vitro in tissue culture medium. The tissue culture material was multiplied by suc

pact argilous soils. Another major characteristic exhibited by

vest per annual cycle.

Table 1 depicts the glycoside and Reb A percentages for the ‘Criola’ lines described above.

The sweetest of these glycosides is rebaudioside A (Reb A).

35

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 ?eld in Woodland, Calif. between March and April of year 2. The plantlets were then allowed to ?ower, which occurred between July and August of year 2. InAugust of year 2, anthers from the ?owered plantlet were collected before anthesis. Approximately 200 anthers were

US PP22,593 P3 3

4

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 Slevia lines required that the chromosome count of each haploid line be doubled, since haploid lines cannot survive as plants. The full complement of

The HPLC data shoWs that the average percentage of Reb A to Glycoside in Slevia 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 Slevia variety ‘T60’ is a perennial herb With ?liform deep roots. The ‘T60’ variety has a long stem that is on average 65-100 cm tall, erect, hairy, and slender. The stem

chromosomes Was regenerated by using successive colchi cine 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 trans ferred into a hormone-free medium for multiplication. Mul tiplication of the diploid plantlets resulted in the creation of a

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

unique line of plants With unique characteristics distinguish able forrn the original four ‘Criola’ lines. These neW Slevia

annually.

lines Were named T-lines. The T-lines Were groWn out in 2

The stem of the ‘T60’ variety produces an average of 3 to 7 lateral branches. The average length of each lateral branches

inch plugs, then transferred to 4 inch pots, and then planted in the ?eld 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

20

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 25

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

3 cm and an average Width of 0.6 to 1 cm. The blades are apex

30

Of the neW Slevia T-lines, three varieties Were selected that

the leaf blades are olive-green to broWnish green in color 35

(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 corym bose in?orescences at the terminal ends of the branches, and 40

produced high levels of Reb A. The production of high levels

have a peduncle that is on average 1-4 cm long, and very slender. Pedicels of each capitulum are slender and on aver age 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

of Reb A in these Slevia T-lines Will alloW large amounts of Reb A to be harvested. The present Slevia variety ‘T60’ is one

142A), and the upper half has a yelloWish (RHS 142C) color.

of the three selected T-lines developed by the methods described herein. The Slevia variety ‘T60’ Was derived from the T-line ‘T6’. The ‘T6’ line Was derived from ‘Criola’ line ‘IAN-135’.

nent 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,

All neW T-lines Were groWn out in greenhouses in Wood

land, Calif. for planting in the ?eld, to alloW for further testing and breeding for seed.

and sub sessile. The intemodes 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

obtuse, base cuneate, margins entire serate on the upper half, three primary veins arise from the leaf base raised and promi

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 Slevia plants Were as high as 89.2% of total glycoside levels. After the HPLC data Was obtained, the T-line Slevia plants Were planted in the ?eld in Woodland, Calif., and groWn to maturity. At maturity, the production rates of glycosides Were evaluated.

ranges from 30 to 40 cm. The average diameter of the lateral branches ranges from 5 to 7 mm. The lateral branches are light

The stems at maturity are broWn-reddi sh in color (RHS 35A 45

35B). The Slevia variety ‘T60’ has 5 phyllaries that are ?nely

hairy, green (RHS 129B) When fresh, linear to subulate, 4-5

Table 2 depicts HPLC data on the percentage of glycoside and Reb A for samples of the Slevia variety ‘T60’.

mm long, and acute to rounded at apex. Each capitulum is 50

TABLE 2

made up to 5 disk ?orets greenish White (RHS 155C). The seeds are achenes bearing numerous, equally long pappus aWns.

The Slevia variety ‘T60’ expresses high concentrations of Samples of mature

Total

Total

Reb A to

‘T 60’ Slevia plants

Glycoside

Reb A

Glycoside

09-0144 09-0154 09-0165 09-0166

15.18 16.96 14.58 15.05

12.58 14.39 12.00 12.46

82.8% 84.8% 82.3% 82.8%

total glycosides, With a total of rebaudioside A concentration of greater than 80%. The Slevia variety ‘T60’ has an average 55

biological cycle of 90-115 days. We claim:

1. A novel and distinct variety of Slevia plant named ‘T60’ having the characteristics described and illustrated herein. *

*

*

*

*

US. Patent

Mar. 20, 2012

US PP22,593 P3

PEG 3.

Stevia plant named 'T60'

Apr 28, 2010 - plugs and then into 4 inch pots between December of year 1 and January of ... levels of Reb A in these neW T-line Slevia plants Were as high.

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