Optimized Method for Cultivation and Microbial Bioaugmentation of Typha latifolia (Cattail)

Summary

Typha latifolia, which primarily propagates asexually through rhizomes, poses collection challenges due to its extensive root system. This paper presents a method for growing T. latifolia from seed, facilitating easier lab cultivation and offering the potential for sterile plant growth and early microbial bioaugmentation.

Abstract

Typha latifolia, more commonly known as the broadleaf cattail or the common bulrush, has a globally reaching range and dominates wetland ecosystems in North America. While different species of cattail are often considered invasive in North America, T. latifolia is considered the native species to the region and is found throughout the entire continent as the dominant Typha species. Historically, Typha has served various functions, from food sources to building materials. More recently, T. latifolia has emerged as a prominent species to aid bioremediation efforts. With increasing interest in the development of constructed wetland treatment systems (CWTS) for contaminant remediation, reproducible techniques to cultivate cattail in a laboratory environment are necessary. The work presented here examined and tested various growth parameters for the successful cultivation of T. latifolia from seed. Successful germination of Typha species involves scarification (rupture of the seed coat), which was achieved using mechanical techniques for large-scale production. Early seed establishment was shown to favor low nutrient growth conditions for the first week, followed by the introduction of fertilizer in subsequent weeks to enhance post-transplant survival. For microbial bioaugmentation of the plant system, results showed that soaking the seeds in inoculum leads to more extensive colonization of the root tissue and long-term bacterial persistence. An optimized seed sterilization technique using a combination of bleach and detergent was used to improve microorganism colonization success. The growth vessels, both sterile and non-sterile, designed in this study support the long-term growth of T. latifolia under various conditions.

Introduction

As a cost and energy-efficient technology, constructed wetland treatment systems (CWTS) have gained significant popularity for the remediation of environmental contaminants^1,2,3. CWTS uses physical, chemical, and biological processes to remove, transform, or stabilize contaminants. While the physical and chemical processes involved in chemical turnover have been well characterized since the genesis of CWTS, the impact of vegetation remained largely enigmatic². In recent years, there has been an increased focus on understanding the mechanisms by which p....

Protocol

1. Seed scarification

1. Cattail seeds were sourced from a wetland in Calgary, AB, Canada, in fall 2023 (51.11312° N, 114.39381° W). Collect cattail inflorescence in the fall and store in a paper bag in the dark until use. If still on the stalk, collect the inflorescence in the spring, but seed recovery will be diminished.
2. Using garden shears, cut the plant stem ~2 cm from the base of the inflorescence. Pull the seeds off the inflorescence and place into a laboratory blender until the volume of the blender is approximately one-quarter full. This is approximately 250 mL of seeds not compacted.
3. Fill the blende....

Results

Figure 1 shows a representation of a Typha seed that has gone through the scarification process, along with an incompletely scarified seed with the beak removed, and a non-scarified seed. Ensure that the blending time is sufficient to produce primarily fully scarified seeds.

After scarifying and drying the seeds, the viability of the seeds should be tested using the non-sterile germination technique. At the end of 7 days, a minimum of 20% of the seeds sho.......

Discussion

The protocol presented here provides a detailed guide for the growth of cattail species from seed for laboratory applications. While cattail can be easily propagated from rhizome cuttings, starting plants from seed allows for genetic variation within a sample set, ensures plants are at equal growth stages, and provides the opportunity for early colonization for microbial bioaugmentation experiments. It is important to correctly identify the species being collected, as propagation of more invasive cattail species may prom.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
0.2 µm sterile filter	VWR	514-4126
3′,5′-Dimethoxy-4′-hydroxyacetophenone	PhytoTech Labs	S7777
Culture boxes with lids (77 mm × 77 mm × 97 mm)	Millipore Sigma	V8505
Heat-resistant silicone sealant hi-temperature	Imperial Manufacturing Group	KK0205
Laboratory sealing film	Millipore Sigma	P7793
Murashige and Skoog media	PhytoTech Labs	M401
Organosilicone surfactant	PhytoTech Labs	S7777
Phytoagar	GoldBiotechnology	P1003
Polysorbate 20	Roche	11332465001