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Method for carrying out callus induction and differentiation and growth of seedlings on Typha latifolia root tips

A callus and callus induction technology, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of small seeds, difficult shelling and separation of embryos

Active Publication Date: 2012-07-18
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reason is that the seeds of the broad-leaved cattail are too small to shell and separate the embryos.

Method used

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  • Method for carrying out callus induction and differentiation and growth of seedlings on Typha latifolia root tips
  • Method for carrying out callus induction and differentiation and growth of seedlings on Typha latifolia root tips
  • Method for carrying out callus induction and differentiation and growth of seedlings on Typha latifolia root tips

Examples

Experimental program
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Effect test

Embodiment 1

[0026] A method for inducing callus and differentiating into shoots from the root tips of Typha broadleaf, the method comprising the steps of:

[0027] (1) dry, sterilize, germinate and take root of the seeds of Typha broadleaf;

[0028] The tidbits of cattail broadleaves were collected from Yangshan Park in the Xianlin area of ​​Nanjing. The method of seed isolation can be found in the reference (Rogers et al., 1998).

[0029] The described drying condition is drying at 42°C for 24 hours.

[0030] Described disinfection comprises the steps:

[0031] (1a) Soak the seeds twice in sterilized distilled water for 5 minutes each time, and drain the water;

[0032] (1b) soak the seeds treated in step (1a) with 75% (v / v) ethanol solution for 90 seconds, and pour off the ethanol solution;

[0033] (1c) Add 2.5% (w / w) sodium hypochlorite solution to the treated seeds in step (1b) for disinfection for 15 minutes, shake constantly during disinfection, repeat once, and pour off the so...

Embodiment 2

[0041] Embodiment 2: test method and statistical analysis of data.

[0042] Nine-week-old calli were dried at 42°C for 2 days and then weighed. Since the callus is small, in order to avoid errors, every 5 callus was counted as one portion when weighing. Table 2 is the weight of callus. When the callus induced seedlings for 3 weeks, the number of seedlings emerging from each piece of callus was recorded. The length of the shoots after 9 weeks was measured. All the above determinations were set in triplicate. Data were analyzed using ANOVA and Tukey's test. The confidence interval is 95%. The analysis software is R.

Embodiment 3

[0043] Example 3: Establishment of callus induction method.

[0044] The sterilized seeds were added with 30g L -1 sucrose, pH 5.8 MS culture medium, all can germinate under continuous light, but less than 1% of the seeds can germinate under dark conditions, and it is difficult to form callus on the callus induction culture machine organize. Experiments showed that 3-day-old shoots had a good success rate of callus induction. The 2-day and 3-day-old buds are better than the 1-day-old seedlings for root induction. As the shoots grow, the efficiency of callus induction will decrease. In summary, 2-day-old shoots were used to induce roots. More than 90% of the shoots on the rooting medium produced healthy roots. After rooting and culturing for 5-7 days, a 2-3mm root tip was cut out and transferred to a callus induction medium. The experiment found that no callus could be induced on the medium without adding plant hormones, and in the control experiment, the efficiency of 2,...

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Abstract

The invention discloses a method for carrying out callus induction and differentiation and growth of seedlings on Typha latifolia root tips. The method comprises the following steps of: drying, sterilizing, germinating and rooting Typha latifolia seeds; carrying out callus induction and differentiation and growth of seedling on the rooted Typha latifolia root tips, wherein the step comprises the following specific steps of: acquiring 2-3mm root tips from rooted shoot roots, and grafting the root tips to a callus induction induced culture medium and culturing for 9 weeks under the dark condition; grafting the callus onto the callus differentiation culture medium, and carrying out differentiation culture for 4 weeks at 25 DEG C under the continuous illumination condition; grafting seedlingsobtained after differentiation culture onto a rooting culture medium and culturing for 4 weeks; and transferring the seedling subjected to rooting culture into a small flowerpot containing a flower fertilizer, placing the seedlings into a 25-DEG C greenhouse for growth, covering a plastic film above the seedlings at the first week for preventing water from evaporating, watering once every three days, culturing for 40 days, then transferring the small flowerpot to a larger flowerpot, and maintaining water to be 3-5cm.

Description

technical field [0001] The invention relates to an efficient method for inducing callus from the root tip of cattail broadleaves and differentiating it into seedlings. Background technique [0002] Broadleaf cattail is a large plant that occurs naturally in ponds, lakes and swamps. It is widely distributed in tropical and subtropical regions. It has a well-developed root system and an efficient photosynthetic system, which can efficiently utilize non-structural carbohydrates at one time. Therefore, cattails are widely used in natural or artificial wetland sewage treatment systems to treat urban domestic sewage, industrial wastewater, farmland sewage, mining sewage, etc. [0003] What's more important is that Cattail broadleaf has high tolerance to heavy metals such as cadmium, arsenic, zinc and lead. These heavy metals can be enriched in the roots of cattails without causing obvious damage. Some researchers removed heavy metal pollution by rapidly cultivating the roots o...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 杨柳燕杜宏伟武俊刘露肖琳吕志刚许超张全兴
Owner NANJING UNIV
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