Paper · 2020 · Digital Scholarship - UNLV (University of Nevada Reno)

The Role of OsWRKY71 and Its Interacting Proteins in Seed Germination and Early Growth of Cereal Grains

Shin, Margaret Ja

12
claims
24
verbatim quotes
1
citations
3
research areas
✓
Open access

Abstract

During seed germination and early seedling growth, complex molecular and physiological events occur in rice (Oryza sativa) and other cereal grains. As the seed transitions to vegetative tissue, it responds to both favorable and unfavorable environmental conditions and is vulnerable to attack by predation and disease. Although seeds are relatively small and tender in size, extensive and sophisticated molecular networks enables the immobile seed to grow, survive and adapt in its environment. One of the networks I am interested in is in the crosstalk between the gibberellin (GA) and abscisic acid (ABA) signaling pathways. These pathways are interesting because they are largely antagonistic. GA is a hormone that generally promotes germination and growth- related processes while ABA, also a hormone, promotes seed dormancy and represses growth. Although a great deal of research has been dedicated towards understanding these two pathways, the actual mechanism of crosstalk during seed germination is less understood. Any deficiencies in GA and ABA regulation and response may result in altered interpretation of environmental signals and aberrations in seed development and germination, leading to lower grain yields. My research is dedicated towards deciphering the specific role of Oryza sativa WRKY71 (OsWRKY71; amino acid W-R-K-Y) and harpin-induced1-like (HIL) members in the crosstalk between GA and ABA in rice, with the goal that this research will be used to improve cereal grain yield in areas of the world with limited plant productivity. The crosstalk between GA and ABA directs the synthesis of ?-amylase, which is an enzyme that breaks down starch in seeds to provide energy for germination. OsWRKY71 was shown to be a transcriptional regulator of ?-amylase and was regulated by both GA and ABA in barley. In this study, I have provided a model of the regulation of OsWRKY71 in seed germination in rice. Although it was previously determined that OsWRKY71 negatively regulated ?-amylase, I show that it positively regulated not only germination but also root growth. To support this, I performed seed germination and root elongation assays using knockout mutants of OsWRKY71. Mutant analysis determined that germination in oswrky71 was delayed for approximately 1 day and was able to recover from the delay. Additionally, after 4 days, oswrky71 seedling roots were nearly 2 cm shorter than wildtype (wt), suggesting that OsWRKY71 may regulate other aspects of plant development. This is further supported by analysis of ?-Glucuronidase (GUS) reporter expression of OsWRKY71p-GUS, which indicated that OsWRKY71 was localized to the third node of rice culms. Thus, the function of OsWRKY71 appears to be more complex and versatile than predicted. To further understand the mechanism of OsWRKY71 regulation in rice seed germination, I investigated the role of one of its interacting partners, Oryza sativa harpin-induced1-like 58 (OsHIL58). Using rice aleurone RNA-sequencing data, I found that OsHIL58 was induced upon ABA treatment. Thus, the two proteins may interact during ABA induction. I also annotated the HIL family using in silico methods and identified several other HIL members that were differentially and significantly expressed in the aleurone. One member, OsHIL16, was highly expressed and also coexpressed with an ABA receptor, regulatory component of ABA receptor 9 (RCAR9). Surprisingly, both were repressed by ABA, suggesting that they be involved in the same pathway in the aleurone aside from OsWRKY71 regulation. From this annotation, I also identified and compiled a large family of 104 unique HIL members expressed in various rice tissues. A classification system was designed based on the presence of several conserved amino acid motifs: NPN, RPP, and YQYF. Most HIL members, including OsHIL16 and -58, were Group I members with all three motifs present. These and further analyses suggest that HILs may have multiple roles in plant development, including in seed germination.

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Claims 12

Mechanismobservedoriginal resultpositivemeasured

Claim · model-written summary

OsWRKY71 positively regulated rice germination.

Gene
OsWRKY71t0
Outcome
germinatione0

Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Oryza sativa
Oryza sativa · Taxon · picture from its Wikimedia Commons categoryDesmanthus4food · CC BY-SA 3.0 us · Wikimedia Commons ↗

Verbatim

e0

“I show that it positively regulated not only germination but also root growth.”

✓ located verbatim in the source · abstract, characters 2009–2087 · show in text ↑
t0

“OsWRKY71 and Its Interacting Proteins in Seed Germination and Early Growth of Cereal Grains”

✓ located verbatim in the source · title, characters 12–103 · show in text ↑
e1

“rice (Oryza sativa)”

✓ located verbatim in the source · abstract, characters 103–122 · show in text ↑
Mechanismobservedoriginal resultpositivemeasured

Claim · model-written summary

OsWRKY71 positively regulated rice root growth.

Gene
OsWRKY71t0
Outcome
root growthe0

Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Oryza sativa
Oryza sativa · Taxon · picture from its Wikimedia Commons categoryDesmanthus4food · CC BY-SA 3.0 us · Wikimedia Commons ↗

Verbatim

e0

“I show that it positively regulated not only germination but also root growth.”

✓ located verbatim in the source · abstract, characters 2009–2087 · show in text ↑
t0

“OsWRKY71 and Its Interacting Proteins in Seed Germination and Early Growth of Cereal Grains”

✓ located verbatim in the source · title, characters 12–103 · show in text ↑
e1

“rice (Oryza sativa)”

✓ located verbatim in the source · abstract, characters 103–122 · show in text ↑
Observationobservedoriginal resultnegativemeasured

Claim · model-written summary

oswrky71 mutant germination was delayed about one day and recovered.

Material
oswrky71e0
Outcome
germination in oswrky71e0
Direction
delayede0
Value
approximately 1 daye0

Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Oryza sativa
Oryza sativa · Taxon · picture from its Wikimedia Commons categoryDesmanthus4food · CC BY-SA 3.0 us · Wikimedia Commons ↗

Verbatim

e0

“germination in oswrky71 was delayed for approximately 1 day and was able to recover from the delay.”

✓ located verbatim in the source · abstract, characters 2229–2328 · show in text ↑
t0

“rice (Oryza sativa)”

✓ located verbatim in the source · abstract, characters 103–122 · show in text ↑
Observationobservedoriginal resultnegativemeasured

Claim · model-written summary

At four days, oswrky71 roots were nearly 2 cm shorter than wild type roots.

Material
oswrky71e0
Outcome
oswrky71 seedling rootse0
Direction
shortere0
Value
nearly 2 cme0
Comparator
wildtype (wt)e0

Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Oryza sativa
Oryza sativa · Taxon · picture from its Wikimedia Commons categoryDesmanthus4food · CC BY-SA 3.0 us · Wikimedia Commons ↗

Verbatim

e0

“after 4 days, oswrky71 seedling roots were nearly 2 cm shorter than wildtype (wt)”

✓ located verbatim in the source · abstract, characters 2343–2424 · show in text ↑
t0

“rice (Oryza sativa)”

✓ located verbatim in the source · abstract, characters 103–122 · show in text ↑
Observationobservedoriginal resultmeasured

Claim · model-written summary

OsWRKY71 reporter expression localized at the third node of rice culms.

Organ / stage
third node of rice culmse0
Gene
OsWRKY71e0
Outcome
OsWRKY71 was localized to the third node of rice culmse0

Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Oryza sativa
Oryza sativa · Taxon · picture from its Wikimedia Commons categoryDesmanthus4food · CC BY-SA 3.0 us · Wikimedia Commons ↗

Verbatim

e0

“OsWRKY71 was localized to the third node of rice culms.”

✓ located verbatim in the source · abstract, characters 2622–2677 · show in text ↑
Intervention effectobservedoriginal resultpositivemeasured

Claim · model-written summary

ABA treatment induced OsHIL58 in rice aleurone RNA sequencing data.

Treatment
ABA treatmente0
Gene
OsHIL58e0
Outcome
OsHIL58 was inducede0
Direction
inducede0

Not reportedComparator

Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Oryza sativa
Oryza sativa · Taxon · picture from its Wikimedia Commons categoryDesmanthus4food · CC BY-SA 3.0 us · Wikimedia Commons ↗

Verbatim

e0

“Using rice aleurone RNA-sequencing data, I found that OsHIL58 was induced upon ABA treatment.”

✓ located verbatim in the source · abstract, characters 2959–3052 · show in text ↑
Observationobservedoriginal resultsignificant (as stated)measured

Claim · model-written summary

Several HIL members were differentially and significantly expressed in aleurone.

Organ / stage
aleuronee0
Gene
HILe0
Outcome
HIL members that were differentially and significantly expressede0

Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Oryza sativa
Oryza sativa · Taxon · picture from its Wikimedia Commons categoryDesmanthus4food · CC BY-SA 3.0 us · Wikimedia Commons ↗

Verbatim

e0

“identified several other HIL members that were differentially and significantly expressed in the aleurone.”

✓ located verbatim in the source · abstract, characters 3171–3277 · show in text ↑
t0

“rice (Oryza sativa)”

✓ located verbatim in the source · abstract, characters 103–122 · show in text ↑
Observationobservedoriginal resultmeasured

Claim · model-written summary

OsHIL16 was highly expressed in aleurone.

Gene
OsHIL16e0
Outcome
OsHIL16, was highly expressede0

Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Oryza sativa
Oryza sativa · Taxon · picture from its Wikimedia Commons categoryDesmanthus4food · CC BY-SA 3.0 us · Wikimedia Commons ↗

Verbatim

e0

“One member, OsHIL16, was highly expressed and also coexpressed with an ABA receptor, regulatory component of ABA receptor 9 (RCAR9).”

✓ located verbatim in the source · abstract, characters 3278–3410 · show in text ↑
t0

“rice (Oryza sativa)”

✓ located verbatim in the source · abstract, characters 103–122 · show in text ↑
Associationobservedoriginal resultmeasured

Claim · model-written summary

OsHIL16 was coexpressed with RCAR9.

Gene
OsHIL16e0
Outcome
coexpressed with an ABA receptor, regulatory component of ABA receptor 9 (RCAR9)e0

Not reportedDirection

Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Oryza sativa
Oryza sativa · Taxon · picture from its Wikimedia Commons categoryDesmanthus4food · CC BY-SA 3.0 us · Wikimedia Commons ↗

Verbatim

e0

“OsHIL16, was highly expressed and also coexpressed with an ABA receptor, regulatory component of ABA receptor 9 (RCAR9).”

✓ located verbatim in the source · abstract, characters 3290–3410 · show in text ↑
t0

“rice (Oryza sativa)”

✓ located verbatim in the source · abstract, characters 103–122 · show in text ↑
Intervention effectobservedoriginal resultnegativemeasured

Claim · model-written summary

ABA repressed both OsHIL16 and RCAR9.

Treatment
ABAe0
Outcome
repressede0
Direction
repressede0

Not reportedGeneComparator

Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Oryza sativa
Oryza sativa · Taxon · picture from its Wikimedia Commons categoryDesmanthus4food · CC BY-SA 3.0 us · Wikimedia Commons ↗

Verbatim

e0

“both were repressed by ABA, suggesting that they be involved in the same pathway in the aleurone aside from OsWRKY71 regulation.”

✓ located verbatim in the source · abstract, characters 3425–3553 · show in text ↑
t0

“rice (Oryza sativa)”

✓ located verbatim in the source · abstract, characters 103–122 · show in text ↑
Method / resourceobservedoriginal resultbasis n/a

Claim · model-written summary

A family of 104 unique HIL members was compiled across rice tissues.

Gene
HILe0
Outcome
HIL members expressed in various rice tissuese0
Value
104e0

Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Oryza sativa
Oryza sativa · Taxon · picture from its Wikimedia Commons categoryDesmanthus4food · CC BY-SA 3.0 us · Wikimedia Commons ↗

Verbatim

e0

“identified and compiled a large family of 104 unique HIL members expressed in various rice tissues.”

✓ located verbatim in the source · abstract, characters 3583–3682 · show in text ↑
t0

“rice (Oryza sativa)”

✓ located verbatim in the source · abstract, characters 103–122 · show in text ↑
Observationobservedoriginal resultmeasured

Claim · model-written summary

Most HIL members belonged to Group I with NPN, RPP and YQYF motifs.

Gene
HILe0
Outcome
Group I members with all three motifs presente0

Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Oryza sativa
Oryza sativa · Taxon · picture from its Wikimedia Commons categoryDesmanthus4food · CC BY-SA 3.0 us · Wikimedia Commons ↗

Verbatim

e0

“Most HIL members, including OsHIL16 and -58, were Group I members with all three motifs present.”

✓ located verbatim in the source · abstract, characters 3802–3898 · show in text ↑
t0

“rice (Oryza sativa)”

✓ located verbatim in the source · abstract, characters 103–122 · show in text ↑

(+)-abscisic acid · organism · Oryza sativa (rice)Positive and negative results

7 papers: 5 positive, 2 negative or no effect

4 papers in the same direction as this paper · 2 in the opposite direction

  1. no effect
  2. negative
  3. positive

3 papers more in this group

See all 7 papers →

Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.