rpc00050-YG1-3.2 (2017.10.19; revised 2024.1.29)
RIKEN BRC plant cell line documentation

rpc00050

Arabidopsis thaliana YG1 cell suspension cultureComponents• Domestic delivery: A 50-mL plastic conical centrifuge tube, containing cellsuspension• Overseas delivery: A 250-mL plastic Erlenmeyer ﬂask, containing cells
placed on semi-solid medium
Notice• Subculture the cells to fresh medium immediately after arrival [Notes I, II].
• Do not store the cell culture in a refrigerator and a freezer.
• Maintain aseptic conditions of the cell culture, and work in a laminar ﬂowcabinet.
Method• Culture medium: mLS medium, 0.2 mg/L 2,4-D, pH 5.8 (medium no. 1)[Materials III]
• Culture conditions: 27℃, dark, 130 rpm [Methods II]• Subculture: 7-day intervals [Methods I]
Citation of cell line
When results obtained by using this cell line are published in a scientiﬁc journal, it should be cited in the following manner: “Arabidopsis thaliana YG1cell line (rpc00050) was provided by the RIKEN BRC through the National
BioResource Project of the MEXT, Japan.”Experimental Plant Division
RIKEN BioResource Research Center (BRC)
Koyadai 3-1-1, Tsukuba, Ibaraki 305-0074
JapanFAX: +81 29 836 9053
E-mail: plant.brc@riken.jp
http://epd.brc.riken.jp/en/

1

Introduction
Arabidopsis YG1 cell line was established from a seedling of Arabidopsis thaliana (L.)
Heynh. accession Columbia (Nishikiori et al. 2011, Yoshikawa et al. 2021). The YG1cells are grown in a modiﬁed Linsmaier and Skoog (mLS) medium supplemented with
0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), pH 5.8. Our YG1 cell culture has been
maintained in the dark at 27℃ with rotary shaking at 130 rpm and subcultured at 7-day
intervals.

Materials
Chemicals and stock solutions(All stock solutions are stored at 4℃)
A) MS salt mix
Murashige and Skoog Plant Salt Mixture, FUJIFILM Wako Pure Chemical
Corporation (#392-00591)
B) SucroseC) BY2_P
KH2 PO480 mg/mLD) LS_VT_modiﬁed
Thiamine·HCl
myo-Inositol

0.4 mg/mL
40 mg/mLE) 2,4-D (0.2 mg/mL)
2,4-D sodium monohydrate
0.236 mg/mL(2,4-Dichlorophenoxy)acetic acid sodium salt monohydrate, Sigma-Aldrich(D6679)F) KOH (1 N)
GlasswareA) Erlenmeyer ﬂask (300 mL), capped with two layers of aluminum foilB) Pipette (10 mL; large tip opening) and a bulb, sterilized by autoclaving at 121℃ for
20 min2Preparation of mLS medium (medium no. 1)
1. Dissolve the following chemicals in approximately 800 mL of distilled water.
MS salt mix
Sucrose1 bag (1 L)
30 g

2. Add following stock solutions, and ﬁll up to approximately 950 mL with distilledwater.
BY2_P
LS_VT_modiﬁed2,4-D (0.2 mg/mL)

2.5 mL
2.5 mL
1 mL

3. Adjust the pH of the solution to 5.8 with KOH (1 N), and ﬁll up to 1 L with distilledwater.
4. Pour 80 mL of the medium into a 300-mL ﬂask.
5. Autoclave the ﬂask at 121℃ for 20 min.

Methods
1. Leave to stand a ﬂask containing a 7-day-old culture until the cells settle to the bottom of the ﬂask.
2. Transfer 4–6 mL of the sedimented cells to 80 mL of fresh mLS medium with apipette.
3. Incubate cell cultures on a rotary shaker at 130 rpm under the dark condition at27℃.

Notes• For domestic customers: We send YG1 cell suspension in a 50-mL disposable conical centrifuge tube. The cells should be transferred to fresh mLS medium immediately after arrival.
• For overseas customers: We send YG1 cells placed on semi-solid mLS medium ina 250-mL disposable Erlenmeyer ﬂask. The cells should be transferred to freshmLS medium immediately after arrival. Collect the cells from the semi-solidmedium with a spatula and transfer them to Erlenmeyer ﬂasks containing fresh liquidmedium.
• In order to maintain YG1 cell suspension cultures stably, it is essential to transfer anadequate amount of cells to fresh mLS medium in every subculture. The amount of3cells may vary from one lab to another, because proliferation of YG1 cells is aﬀectedby culture conditions, such as a room temperature, rotation speed of a rotary shaker,
and aeration condition of the culture.
• In order to obtain good aeration of a suspension culture, a silicone sponge plug
may be used instead of the aluminum foil cap (e.g., cap-type Silicosen; Shin-Etsu
Polymer, Tokyo, Japan; https://www.shinpoly.co.jp/en/product/product/medical/plugs.html).

References
Nishikiori M, Mori M, Dohi K, Okamura H, Katoh E, Naito S, Meshi T, Ishikawa M (2011)
A host small GTP-binding protein ARL8 plays crucial roles in tobamovirus RNA
replication. PLoS Pathogen 7: e1002409. DOI: 10.1371/journal.ppat.1002409
Yoshikawa M, Han Y-W, Fujii H, Aizawa S, Nishino T, Ishikawa M (2021) Cooperative
recruitment of RDR6 by SGS3 and SDE5 during small interfering RNA ampliﬁcation in Arabidopsis. Proceedings of National Academy of Sciences of USA 118:
e2102885118. DOI: 10.1073/pnas.2102885118

4Appendix A: Formulation of culture medium
Table A.1. modiﬁed Linsmaier and Skoog medium(medium no. 1)
ChemicalConcentration (mg/L)

KNO31900

NH4 NO31650

CaCl2 ·2H2 O

440

MgSO4 ·7H2 O

370

KH2 PO4370

H3 BO3

6.2

MnSO4 ·4H2 O

22.3

ZnSO4 ·7H2 O

8.6

KI0.83Na2 MoO4 ·2H2 O

0.25

CuSO4 ·5H2 O

0.025

CoCl2 ·6H2 O

0.025

FeSO4 ·7H2 O

27.8Na2 -EDTA

37.3

Thiamine·HCl

1

myo-Inositol

100

Sucrose30000

2,4-D sodium monohydrate

0.2365