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Plants having improved growth characteristics and method for making the same

A growth characteristic and plant technology, applied in the field of molecular biology, can solve the problems of not fully reflecting the function of HKT1 and the complex conditions of HKT1 expression downregulation, and achieve the effect of improving growth characteristics and increasing yield

Inactive Publication Date: 2007-12-19
CROPDESIGN NV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as shown by Laurie et al. (Plant J.32, 139-149, 2002), the phenotype of overexpressed HKT1 plants was not much different from that of control plants: under NaCl stress conditions, the fresh weight of HKT1 overexpressed lines decreased to that of control plants. Plants are similar to the degree, and, compared with the control, the Na in the root of the overexpression line + content did not increase significantly
Laurie et al. also proved that the conditions for down-regulation of HKT1 expression are quite complicated
To date, existing technologies related to HKT1 have mainly focused on cation transport and ion homeostasis, and most studies have been performed in yeast or Xenopus oocytes, and these results may not fully reflect HKT1 in plants. Function

Method used

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  • Plants having improved growth characteristics and method for making the same
  • Plants having improved growth characteristics and method for making the same
  • Plants having improved growth characteristics and method for making the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0149] Example 1: Gene cloning

[0150] The Arabidopsis thaliana seedling cDNA library (Invitrogen, Paisley, UK) was used as a template to amplify the Arabidopsis HKT gene (CDS1523) by PCR. After reverse transcription of the RNA extracted from the seedlings, the cDNA was cloned into pCMV Sport 6.0. The average insert size of the library is 1.5kb, and the number of original clones is in the order of 1.59×10 7 cfu. In 6×10 11 After the first amplification of cfu / ml, the original titer is determined to be 9.6×10 5 cfu / ml. After extracting the plasmid, 200 ng template was used in 50 μl PCR mix. The primers used for PCR amplification (which include the AttB site of Gateway recombination) are prm3283 (SEQ ID NO: 5) and prm8443 (SEQ ID NO: 6). PCR was performed using Hifi Taq DNA polymerase under standard conditions. The same standard method was used to amplify and purify the 1613 bp PCR fragment. Then proceed to the first step of the Gateway process, the BP reaction, during which the PC...

Embodiment 2

[0151] Example 2: Vector construction

[0152] Next, an LR reaction was performed using the entry clone p036 and the designated vector p56 for rice transformation. This vector contains the following parts as functional elements within the T-DNA boundary: a plant selectable marker; a visual marker; a Gateway expression cassette of LR designed to recombine in vivo with the target sequence that has been cloned into the clone. The rice WSI18 promoter (PRO0151) for constitutive expression is located upstream of this Gateway box. After the LR recombination step, the resulting expression vector p060 (Figure 4) contains the expression cassette of SEQ ID NO: 4, which is transformed into Agrobacterium strain LBA4044, and then into rice plants. The transformed rice plants were grown, and then the parameters described in Example 3 were tested.

Embodiment 3

[0153] Example 3: Conversion evaluation

[0154]Approximately 15 to 20 independent TO transformants were produced. The primary transformants were transferred from the tissue culture room to the greenhouse for growth and T1 seeds were harvested. Five events were retained, among which a 3:1 separation of the presence / absence of the transgene occurred in the T1 progeny. Through visual marker screening, 10 T1 seedlings containing the transgene (heterozygous and homozygous) and 10 T1 seedlings lacking the transgene (null zygote) were selected in each event. Transfer the selected T1 plants to the greenhouse. Each plant is given a unique bar mark to clearly associate the phenotypic data with the corresponding plant. The selected T1 plants are grown in the soil of a 10cm diameter flowerpot under the following environmental conditions: photoperiod = 11.5 hours, daylight intensity = 30,000 lux or more, day temperature = 28°C or higher, night temperature = 22°C, Relative humidity = 60-70%. T...

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Abstract

The present invention concerns a method for improving growth characteristics of plants by increasing activity in a plant of an HKT protein or a homologue thereof. One such method comprises introducing into a plant an HKT nucleic acid molecule or functional variant thereof. The invention also relates to transgenic plants having improved growth characteristics, which plants have increased expression of a nucleic acid encoding an HKT protein. The present invention also concerns constructs useful in the methods of the invention.

Description

Technical field [0001] The present invention generally relates to the field of molecular biology and relates to methods of improving plant growth characteristics. More specifically, the present invention relates to by increasing Na in plants + -K + The method of cotransporter (HKT) or its homologue activity to improve plant growth characteristics, especially yield. The present invention also relates to plants with increased HKT activity, which have improved growth characteristics relative to corresponding wild-type plants. The invention also provides constructs useful in the methods of the invention. technical background [0002] In view of the ever-increasing world population and the shrinking area of ​​agricultural arable land, improving agricultural efficiency and increasing the diversity of horticultural plants have been the main goals of agricultural research. The conventional method of crop and horticultural improvement is to use selective breeding techniques to identify pl...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/29
CPCC12N15/8261Y02A40/146
Inventor A·I·桑兹莫林纳罗
Owner CROPDESIGN NV
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