Os04g0497200 - Revision history

Xysj2012: /* Expression */

2016-09-19T08:47:24Z

‎Expression

Xysj2012: /* Mutation */

2016-09-19T08:47:04Z

‎Mutation

Liman at 14:37, 18 September 2016

2016-09-18T14:37:49Z

Liman: /* References */

2016-09-18T14:36:48Z

‎References

Liman at 14:36, 18 September 2016

2016-09-18T14:36:34Z

Liman: /* Subcellular localization */

2016-09-18T14:33:52Z

‎Subcellular localization

Liman: /* Mutation */

2016-09-18T14:26:35Z

‎Mutation

Liman: /* Mutation */

2016-09-18T14:15:20Z

‎Mutation

Liman: /* Mutation */

2016-09-18T14:12:33Z

‎Mutation

Liman: /* Annotated Information */

2016-09-18T14:09:16Z

‎Annotated Information

@@ Line 33: / Line 33: @@
 * Since the OsGLU3 is involved in root cell elongation and division, the researchers expected that it was expressed in the elongating root. To check that, we analyzed the relative OsGLU3 transcripts levels in root, stem-base, stem, leaf, and panicle by qRT–PCR. It showed that the OsGLU3 is highly expressed in root tissue and has relatively lower expression in the other tissues (Figure 5)<ref name="ref1" />.
-[[File:Os04g0497200-4.png|center|thumb|400px|'''Figure 5. OsGLU3 Protein Is Strongly Presented in Elongation Region of Root and Lateral Root, Crown Root Primodia. GUS staining in various organs of proOsGLU3:OsGLU3:GUS transgenic plants.''' '' <ref name="ref1" />.'']]
+[[File:Os04g0497200-4.png|right|thumb|400px|'''Figure 5. OsGLU3 Protein Is Strongly Presented in Elongation Region of Root and Lateral Root, Crown Root Primodia. GUS staining in various organs of proOsGLU3:OsGLU3:GUS transgenic plants.''' '' <ref name="ref1" />.'']]
 ===Subcellular localization===

@@ Line 12: / Line 12: @@
-[[File:Os04g0497200-1.png|center|thumb|400px|'''Figure 1. Osglu3-1 Is Defective in Root Cell Elongation and Division.
+[[File:Os04g0497200-1.png|right|thumb|400px|'''Figure 1. Osglu3-1 Is Defective in Root Cell Elongation and Division.
 (A) Seedlings of wild-type (left), Osglu3-1 (middle), and Osglu3-1 transformed with whole OsGLU3 gene (right). Bar = 2 cm.
 Bottom figures are enlargements of the lateral roots in rectangles. Bar = 200 lm.

@@ Line 4: / Line 4: @@
 ===Function===
-* '''''OsGLU3''''' modulates root cell wall cellulose synthesis and affects root cell elongation and division.
+* '''''OsGLU3''''' modulates root cell wall cellulose synthesis and affects root cell elongation and division<ref name="ref1" />.
-* The researchers also found that phosphate starvation, an environmental stress, altered cell wall cellulose content to modulate root elongation in a '''''OsGLU3'''''-dependant way.
+* The researchers also found that phosphate starvation, an environmental stress, altered cell wall cellulose content to modulate root elongation in a '''''OsGLU3'''''-dependant way<ref name="ref1" />.
 ===Mutation===
-* A rice mutant with short root and short lateral root was obtained at 10 d after germination (DAG) (Figure 1A). The mutant was named Osglu3-1 after the mutation, which will be described later. To determine whether the short root phenotype of Osglu3-1 is due to defects in root cell elongation or division, root longitudinal sections for theWT and Osglu3-1 were made. These showed that the cell length of mature epidermal cells was only one-third of those in the WT, while the root hair length of the mutant is similar to that of WT (Figure 1B and 1C and Supplemental Figure 1). However, the root radical pattern organization and meristem of the mutant seems normal. In order to check the root mitotic activity, a mitotic activity reporter, the OsCYCB1,1:GUS, was incorporated into the mutant. The region expressing the OsCYCB1,1:GUS in the root meristem of mutant is about 90% of that in WT (Figure 1D and 1E). Together, these data suggest that the short root phenotype of Osglu3-1 results from defects in both root cell elongation and division, particularly defect in cell elongation.
+* A rice mutant with short root and short lateral root was obtained at 10 d after germination (DAG) (Figure 1A). The mutant was named Osglu3-1 after the mutation, which will be described later. To determine whether the short root phenotype of Osglu3-1 is due to defects in root cell elongation or division, root longitudinal sections for theWT and Osglu3-1 were made. These showed that the cell length of mature epidermal cells was only one-third of those in the WT, while the root hair length of the mutant is similar to that of WT (Figure 1B and 1C and Supplemental Figure 1). However, the root radical pattern organization and meristem of the mutant seems normal. In order to check the root mitotic activity, a mitotic activity reporter, the OsCYCB1,1:GUS, was incorporated into the mutant. The region expressing the OsCYCB1,1:GUS in the root meristem of mutant is about 90% of that in WT (Figure 1D and 1E). Together, these data suggest that the short root phenotype of Osglu3-1 results from defects in both root cell elongation and division, particularly defect in cell elongation<ref name="ref1" />
 [[File:Os04g0497200-1.png|center|thumb|400px|'''Figure 1. Osglu3-1 Is Defective in Root Cell Elongation and Division.
@@ Line 19: / Line 20: @@
 (E) The width of OsCYCB1;1:GUS expression region in Osglu3-1 and WT.''' '' <ref name="ref1" />.'']]
-* To determine whether the '''''OsGLU3''''' participates in cell wall synthesis, the content of the root cell wall components of '''''Osglu3''''' mutants was measured. It was found that Osglu3-1 has 20% less crystalline cellulose in its roots than that in WT (Figure 3C). Then the WT and the Osglu3-1 mutants were grown on a solid media with 3% glucose. The addition of 3% glucose to the culture medium almost rescued the short lateral root defects of Osglu3-1, although the treatment inhibited the primary root elongation in the WT plants, SSBM (Figure 3A). To determine whether the complementation is due to the restoration of cellulose content in the Osglu3-1, the level of crystalline cellulose in the seedlings grownwith glucose and inWTseedlings was measured. These results showed that Osglu3-1 had almost the same concentration of cell wall cellulose asWT grown onmedium with 3% exogenous glucose. Moreover, the cellulose content of WT and Osglu3-1 had statistically increased after the application of 3% glucose (Figure 3C). The root growth response of Osglu3-2 to glucose treatment was also tested. As with the Osglu3-1 mutant, the application of 3% glucose suppressed the short root and short lateral root defect of Osglu3-2 (Figure 3B).
+* To determine whether the '''''OsGLU3''''' participates in cell wall synthesis, the content of the root cell wall components of '''''Osglu3''''' mutants was measured. It was found that Osglu3-1 has 20% less crystalline cellulose in its roots than that in WT (Figure 3C). Then the WT and the Osglu3-1 mutants were grown on a solid media with 3% glucose. The addition of 3% glucose to the culture medium almost rescued the short lateral root defects of Osglu3-1, although the treatment inhibited the primary root elongation in the WT plants, SSBM (Figure 3A). To determine whether the complementation is due to the restoration of cellulose content in the Osglu3-1, the level of crystalline cellulose in the seedlings grownwith glucose and inWTseedlings was measured. These results showed that Osglu3-1 had almost the same concentration of cell wall cellulose asWT grown onmedium with 3% exogenous glucose. Moreover, the cellulose content of WT and Osglu3-1 had statistically increased after the application of 3% glucose (Figure 3C). The root growth response of Osglu3-2 to glucose treatment was also tested. As with the Osglu3-1 mutant, the application of 3% glucose suppressed the short root and short lateral root defect of Osglu3-2 (Figure 3B)<ref name="ref1" />.
 [[File:Os04g0497200-2.png|center|thumb|400px|'''Figure 3. Three Percent Exogenous Glucose Rescued the Defects of Osglu3-1 and Partially Rescued T-DNA Insert Mutant Osglu3-2.''' '' <ref name="ref1" />.'']]
-* Phosphate starvation could induce root elongation in Osglu3-1 (Figure 4A). Under starvation, both the root cell elongation and division of the primary root were almost restored to the WT level (Figure 4C(d), 4C(f), and 4D). The root cellulose content of Osglu3-1 was also increased (Figure 4F). This indicates that the phosphate-starvation-induced root elongation might be independent of '''''OsGLU3'''''. To further test this, the root growth response of the Osglu3-2 loss-of-function mutant under phosphate starvation was also examined. Similarly to the WT SSBM, the primary root growth of WT Dongjin is also induced by phosphate starvation, with an approximate 30% increase in primary root elongation and 15% increase in root cellulose content. However, the phosphate starvation- induced primary root elongation and cellulose-content increase are abolished in Osglu3-2, which suggests that phosphate starvation-induced primary root elongation depends on the activity of OsGLU3 (Figure 4B).
+* Phosphate starvation could induce root elongation in Osglu3-1 (Figure 4A). Under starvation, both the root cell elongation and division of the primary root were almost restored to the WT level (Figure 4C(d), 4C(f), and 4D). The root cellulose content of Osglu3-1 was also increased (Figure 4F). This indicates that the phosphate-starvation-induced root elongation might be independent of '''''OsGLU3'''''. To further test this, the root growth response of the Osglu3-2 loss-of-function mutant under phosphate starvation was also examined. Similarly to the WT SSBM, the primary root growth of WT Dongjin is also induced by phosphate starvation, with an approximate 30% increase in primary root elongation and 15% increase in root cellulose content. However, the phosphate starvation- induced primary root elongation and cellulose-content increase are abolished in Osglu3-2, which suggests that phosphate starvation-induced primary root elongation depends on the activity of OsGLU3 (Figure 4B)<ref name="ref1" />.
-* To further determine whether the phosphate starvation condition induces the expression of '''''OsGLU3''''', a quantitative RT–PCR analysis of '''''OsGLU3''''' was made. It showed that the phosphate starvation could not induce OsGLU3 expression, which indicates that phosphate starvation might regulate OsGLU3 enzyme activity to induce cellulose synthesis for root elongation.
+* To further determine whether the phosphate starvation condition induces the expression of '''''OsGLU3''''', a quantitative RT–PCR analysis of '''''OsGLU3''''' was made. It showed that the phosphate starvation could not induce OsGLU3 expression, which indicates that phosphate starvation might regulate OsGLU3 enzyme activity to induce cellulose synthesis for root elongation<ref name="ref1" />.
 [[File:Os04g0497200-3.png|center|thumb|400px|'''Figure 4. Phosphate Starvation Could Induce Root Elongation in Osglu3-1, But Not in the T-DNA Insert Mutant Osglu3-2.''' '' <ref name="ref1" />.'']]
 ===Expression===
-* Since the OsGLU3 is involved in root cell elongation and division, the researchers expected that it was expressed in the elongating root. To check that, we analyzed the relative OsGLU3 transcripts levels in root, stem-base, stem, leaf, and panicle by qRT–PCR. It showed that the OsGLU3 is highly expressed in root tissue and has relatively lower expression in the other tissues (Figure 5).
+* Since the OsGLU3 is involved in root cell elongation and division, the researchers expected that it was expressed in the elongating root. To check that, we analyzed the relative OsGLU3 transcripts levels in root, stem-base, stem, leaf, and panicle by qRT–PCR. It showed that the OsGLU3 is highly expressed in root tissue and has relatively lower expression in the other tissues (Figure 5)<ref name="ref1" />.
 [[File:Os04g0497200-4.png|center|thumb|400px|'''Figure 5. OsGLU3 Protein Is Strongly Presented in Elongation Region of Root and Lateral Root, Crown Root Primodia. GUS staining in various organs of proOsGLU3:OsGLU3:GUS transgenic plants.''' '' <ref name="ref1" />.'']]
 ===Subcellular localization===
-* To follow the subcellular distribution of '''''OsGLU3''''', the researchers also generated transgenic plants with OsGLU3–GFP driven by the native OsGLU3 promoter in the Osglu3-1 mutant. The transgenic plants displayed normal root elongation, suggesting that the OsGLU3–GFP protein may reflect the native OsGLU3. In root elongation cells of T1 transgenic seedlings, the GFP fluorescent signals were detected in the plasma member (PM) and intracellular punctuate bodies (Figure 6A). However, the signals were only presented in plasma membrane (Figure 6A and 6B) and in cells within the mature region of the root.
+* To follow the subcellular distribution of '''''OsGLU3''''', the researchers also generated transgenic plants with OsGLU3–GFP driven by the native OsGLU3 promoter in the Osglu3-1 mutant. The transgenic plants displayed normal root elongation, suggesting that the OsGLU3–GFP protein may reflect the native OsGLU3. In root elongation cells of T1 transgenic seedlings, the GFP fluorescent signals were detected in the plasma member (PM) and intracellular punctuate bodies (Figure 6A). However, the signals were only presented in plasma membrane (Figure 6A and 6B) and in cells within the mature region of the root<ref name="ref1" />.
 * When the root was treated with FM4-64 for 30 min, all the compartments of the cell were labeled along the endocytic pathway
-down to the vacuole. These compartments partially co-localized with OsGLU3–GFP in the root meristem, suggesting that the OsGLU3–GFP was located in the endosomal compartment and PM (Figure 6C). In meristem and elongating cells, a 90-min BFA treatment led to the disappearance of the OsGLU3–GFP-labeled punctate structures and a concomitant emergence of large aggregates in root cells (Figure 6E). OsGLU3–GFP largely co-localized with the FM4-64 label to those patches (Figure 6E). This suggest that '''''OsGLU3''''' localizes in the PM and endosomes, and the export of '''''OsGLU3''''' to the PM depends on vesicle transport.
+down to the vacuole. These compartments partially co-localized with OsGLU3–GFP in the root meristem, suggesting that the OsGLU3–GFP was located in the endosomal compartment and PM (Figure 6C). In meristem and elongating cells, a 90-min BFA treatment led to the disappearance of the OsGLU3–GFP-labeled punctate structures and a concomitant emergence of large aggregates in root cells (Figure 6E). OsGLU3–GFP largely co-localized with the FM4-64 label to those patches (Figure 6E). This suggest that '''''OsGLU3''''' localizes in the PM and endosomes, and the export of '''''OsGLU3''''' to the PM depends on vesicle transport<ref name="ref1" />.
 [[File:Os04g0497200-5.png|center|thumb|400px|'''Figure 6. '''''OsGLU3''''' Is Present in PM and Endosomes.

@@ Line 62: / Line 62: @@
 Zhang JW, Xu L, Wu YR, Chen XA, Liu Y, Zhu SH, Ding WN, Wu P, Yi KK. OsGLU3, a putative membrane-bound endo-1,4-beta-glucanase, is required for root cell elongation and division in rice (Oryza sativa L.). Mol Plant. 2012 Jan;5(1):176-86. doi: 10.1093/mp/ssr084. Epub 2011 Oct 5. PubMed PMID: 21976713.
 </ref>
 </references>

@@ Line 30: / Line 30: @@
 ===Expression===
-内切-1,4-β左旋葡聚糖酶在水稻中组织特异性表达，在根尖、侧根和冠状根原基强表达，但在其他组织表达量低，如茎、叶和穗<ref name="ref1"/>。
+* Since the OsGLU3 is involved in root cell elongation and division, the researchers expected that it was expressed in the elongating root. To check that, we analyzed the relative OsGLU3 transcripts levels in root, stem-base, stem, leaf, and panicle by qRT–PCR. It showed that the OsGLU3 is highly expressed in root tissue and has relatively lower expression in the other tissues (Figure 5).
 ===Subcellular localization===
 * To follow the subcellular distribution of '''''OsGLU3''''', the researchers also generated transgenic plants with OsGLU3–GFP driven by the native OsGLU3 promoter in the Osglu3-1 mutant. The transgenic plants displayed normal root elongation, suggesting that the OsGLU3–GFP protein may reflect the native OsGLU3. In root elongation cells of T1 transgenic seedlings, the GFP fluorescent signals were detected in the plasma member (PM) and intracellular punctuate bodies (Figure 6A). However, the signals were only presented in plasma membrane (Figure 6A and 6B) and in cells within the mature region of the root.
 * When the root was treated with FM4-64 for 30 min, all the compartments of the cell were labeled along the endocytic pathway
 down to the vacuole. These compartments partially co-localized with OsGLU3–GFP in the root meristem, suggesting that the OsGLU3–GFP was located in the endosomal compartment and PM (Figure 6C). In meristem and elongating cells, a 90-min BFA treatment led to the disappearance of the OsGLU3–GFP-labeled punctate structures and a concomitant emergence of large aggregates in root cells (Figure 6E). OsGLU3–GFP largely co-localized with the FM4-64 label to those patches (Figure 6E). This suggest that '''''OsGLU3''''' localizes in the PM and endosomes, and the export of '''''OsGLU3''''' to the PM depends on vesicle transport.
 ===Evolution===
-在突变体中，Osglu3-1突变体的根细胞壁中晶体纤维素含量减少，根细胞长度变短，突变体根的分生组织略微变小。外施葡萄糖能够抑制突变体根细胞壁纤维素含量的降低和短根的表型。OsGLU3在不同组织遍在表达，但是在根尖、侧根和冠状根原基强表达。OsGLU3-GFP融合蛋白在根分生组织和伸长区的质膜和FM4-64标记的细胞器表达<ref name="ref1"/>。
 ==Labs working on this gene==

← Older revision		Revision as of 14:33, 18 September 2016
Line 33:		Line 33:

	===Subcellular localization===		===Subcellular localization===
		+	* To follow the subcellular distribution of '''''OsGLU3''''', the researchers also generated transgenic plants with OsGLU3–GFP driven by the native OsGLU3 promoter in the Osglu3-1 mutant. The transgenic plants displayed normal root elongation, suggesting that the OsGLU3–GFP protein may reflect the native OsGLU3. In root elongation cells of T1 transgenic seedlings, the GFP fluorescent signals were detected in the plasma member (PM) and intracellular punctuate bodies (Figure 6A). However, the signals were only presented in plasma membrane (Figure 6A and 6B) and in cells within the mature region of the root.
		+	* When the root was treated with FM4-64 for 30 min, all the compartments of the cell were labeled along the endocytic pathway
		+	down to the vacuole. These compartments partially co-localized with OsGLU3–GFP in the root meristem, suggesting that the OsGLU3–GFP was located in the endosomal compartment and PM (Figure 6C). In meristem and elongating cells, a 90-min BFA treatment led to the disappearance of the OsGLU3–GFP-labeled punctate structures and a concomitant emergence of large aggregates in root cells (Figure 6E). OsGLU3–GFP largely co-localized with the FM4-64 label to those patches (Figure 6E). This suggest that '''''OsGLU3''''' localizes in the PM and endosomes, and the export of '''''OsGLU3''''' to the PM depends on vesicle transport.

	===Evolution===		===Evolution===

← Older revision		Revision as of 14:15, 18 September 2016
Line 22:		Line 22:

	[[File:Os04g0497200-2.png\|center\|thumb\|400px\|'''Figure 3. Three Percent Exogenous Glucose Rescued the Defects of Osglu3-1 and Partially Rescued T-DNA Insert Mutant Osglu3-2.''' '' <ref name="ref1" />.'']]		[[File:Os04g0497200-2.png\|center\|thumb\|400px\|'''Figure 3. Three Percent Exogenous Glucose Rescued the Defects of Osglu3-1 and Partially Rescued T-DNA Insert Mutant Osglu3-2.''' '' <ref name="ref1" />.'']]
		+
		+	* After being grown for 10 d in media without phosphate, the SSBM showed 20% increase of primary root elongation as compared to WT. To determine whether root elongation under phosphate starvation is due to the induction of cell elongation or enhanced mitotic activity, longitudinal sections of the primary root maturation zone were taken and the expression patterns of OsCYCB1;1:GUS under the treatment were analyzed. The longitudinal sections showed that phosphate starvation can lead to an increase of approximately 10% in rice root cell elongation (Figure 4A, 4C(c), and 4D). Also, the starvation can activate root mitotic activity as visualized by a broader GUS staining in the root tip (Figure 4C(e) and 4E). Together, these indicate that phosphate starvation stimulates primary root elongation by inducing root cell elongation and activating root mitotic activity.
		+
		+	[[File:Os04g0497200-3.png\|center\|thumb\|400px\|'''Figure 4. Phosphate Starvation Could Induce Root Elongation in Osglu3-1, But Not in the T-DNA Insert Mutant Osglu3-2.
		+	(A) The phenotype of Osglu3-1 under phosphate starvation. Bar = 2 cm. CK, control; –P, phosphate starvation.
		+	(B) The phenotype of T-DNA insert mutant Osglu3-2 under phosphate starvation. Bar = 2 cm. CK, control; –P, phosphate starvation.
		+	(C) Phosphate starvation induced the cell elongation and mitotic activity in Osglu3-1.
		+	(a, b) The root tip of WT (a) and Osglu3-1 (b) under phosphate starvation. Bar = 1 mm. Left panel, CK; right panel, phosphate starvation.
		+	(c, d) Longitudinal sections of maturation zone ofWT (c) and Osglu3-1 (d) under phosphate starvation. Bar = 50 lm. Left panel, CK; right panel,
		+	phosphate starvation.
		+	(e, f) Expression patterns of OsCYCB1,1:GUS of 10-day-old WT (e) (bar = 100 lm) and Osglu3-1 (f) under phosphate starvation (bar = 50 lm). Left
		+	panel, CK; right panel, phosphate starvation.
		+	(D) Phosphate starvation induced root cell elongation in Osglu3-1.
		+	(E) Phosphate starvation increased the width of OsCYCB1,1:GUS expression region.
		+	(F) The crystalline cellulose (TFA-insoluble) content of Osglu3-1 was partly rescued in phosphate starvation.
		+	(G) The crystalline cellulose (TFA-insoluble) content of Osglu3-2 was not rescued in phosphate starvation. CWM, cell wall material. Data are
		+	mean values from three independent experiments, each performed in triplicate. Error bars represent standard error (SE). Different letters
		+	are used to indicate means that differ significantly (P , 0.05). CWM, cell wall material.''' '' <ref name="ref1" />.'']]

	===Expression===		===Expression===